Systems and methods for reducing fibrosis scarring following tubular organ trauma

Abstract

Aspects of the present disclosure relate to methods and synthetic support structures for regenerating gastrointestinal tissue, such as esophageal tissue.

Description

Technical Field

[0001] This disclosure claims priority to U.S. Provisional Patent Application No. 63 / 394,266, filed on August 1, 2022, the specification of which is incorporated herein by reference.

[0002] The present disclosure relates to engineered tissues that can be used to replace or repair damaged tissue. More specifically, the present disclosure relates to reducing or eliminating stenosis formation following trauma to tubular organs. Background Art

[0003] Engineered biological tissues that can be used to replace or repair damaged tissue are typically prepared by seeding cells on a synthetic structural support and exposing the cells to conditions that allow them to synthesize and secrete extracellular matrix components on the structural support. A variety of techniques have been developed to produce synthetic structural supports, including nanofiber assembly, casting, printing, physical spraying (e.g., using a pump and syringe), electrospinning, electrospraying, and other techniques for depositing one or more natural or synthetic polymers or fibers to form a structural support having a shape and size suitable for transplantation into a subject (e.g., a human subject in need of an organ or engineered tissue area).

[0004] It is estimated that more than 500,000 people are diagnosed with esophageal malignancies each year worldwide. The average incidence of congenital malformations of the esophagus, such as esophageal atresia, is 2.44 per 10,000 births. Chronic esophageal strictures following esophageal injury are also common. Despite advances in minimally invasive resections of early malignancies (eg, endoscopic mucosal resection), the mainstay of treatment for many esophageal diseases remains esophagectomy. Traditionally, an autologous conduit (eg, stomach, small intestine, or colon) is harvested and reintroduced into the chest cavity to restore gastrointestinal continuity. Many children with esophageal atresia or patients who have sustained traumatic or caustic injuries to the esophagus ultimately undergo similar reconstructive procedures. These treatments are often associated with significant morbidity and mortality.

[0005] Autologous catheters have traditionally been used because of the complex structure of the esophagus. The multilayered structure of the esophagus, composed of stratified squamous epithelium, submucosal layer, and external circular and longitudinal muscle layers, serves as a barrier to prevent oral ingesta and contaminants from escaping the gastrointestinal tract. In addition, the combined layers provide a physiological mechanism for propulsion and control the pressure of the food bolus during swallowing or vomiting.

[0006] A variety of conditions can cause damage to tubular organs, ultimately leading to stricture formation. These conditions include injury, disease, and congenital defects that damage the tubular organs and may directly lead to stricture formation or require surgical intervention to restore the integrity and continuity of the organ, which itself may cause stricture formation as a side effect.

[0007] It is desirable to provide structures that can support tissue regeneration and methods for making the same. It is also desirable to provide structures for reducing or eliminating fibrotic scarring in tubular organs, such as gastrointestinal organs (e.g., esophagus, stomach, etc.) and methods for making the same. Also disclosed herein is the use of a cell delivery device in a repair or surgical procedure to promote the orderly generation of tubular tissue during the healing process while reducing the formation of fibrotic scarring. Summary of the invention

[0008] Disclosed herein are the uses of cell delivery devices and support structures in surgery and therapy to reduce the stenosis of hollow tubular organs. Also disclosed herein are the uses of cell delivery devices in surgery to connect hollow tubular organs to another organ. Also disclosed herein are methods for performing surgery and postoperative procedures in hollow tubular organs such as the esophagus, which can reduce or eliminate the structural formation of related organs. Certain embodiments relate to synthetic support structures and related systems capable of producing gastrointestinal tissue (e.g., tissue of the esophagus, stomach, intestines, colon or other hollow gastrointestinal tissues). In some embodiments, the support structure provides guidance for the growth and regeneration of gastrointestinal tract (e.g., esophagus) tissue of the subject. In some embodiments, the regenerated gastrointestinal tissue includes muscle tissue, nervous system tissue, or muscle tissue and nervous system tissue.

[0009] In some embodiments, gastrointestinal tract (e.g., esophagus) tissue regenerates around the support structure. In some embodiments, the support structure is not incorporated into the final regenerated tissue (e.g., the new esophageal tissue does not incorporate the support structure into the regenerated esophageal wall). Therefore, some aspects of the present disclosure relate to guided tissue regeneration, wherein the support structure provides support and / or signals that promote host tissue regeneration without the need to incorporate the support structure into the regenerated tissue (e.g., without the need for the support structure to provide structural or functional support in the final regenerated tissue). Certain embodiments may also include surgical steps that require reduction of fibrotic scar tissue (e.g., areas prone to stenosis).

[0010] In some embodiments, the gastrointestinal tract (eg, esophagus) support structure comprises a biodegradable and / or bioabsorbable material that is absorbed after gastrointestinal tract (eg, esophagus) tissue regeneration has begun (eg, after functional esophageal tissue has been regenerated).

[0011] In some embodiments, a gastrointestinal tract (eg, esophageal) support structure includes one or more structures that can be used to assist in removal of the support structure after gastrointestinal tract (eg, esophageal) tissue regeneration is initiated (eg, after functional esophageal tissue regeneration).

[0012] In some embodiments, the support structure is cellularized with one or more cell types prior to implantation. In some embodiments, the cells are autologous cells. In some embodiments, the cells are progenitor cells or stem cells. In some embodiments, the cells are from bone marrow, adipose tissue, esophageal tissue or other suitable tissues. In some embodiments, the cells can be obtained from various allogeneic sources, including but not limited to sources such as amniotic fluid, umbilical cord blood. In some embodiments, the cells are mesenchymal stem cells (MSC).

[0013] In some embodiments, the support structure is implanted in a site that provides a sufficient stem cell microenvironment (e.g., the esophagus or other gastrointestinal site that provides a stem cell microenvironment) for regenerating tissue in a subject. The term "stem cell microenvironment" as used herein is defined as a microenvironment that interacts with stem cells to regulate the fate of stem cells, including stem cells introduced by the support structure and / or stem cells in the patient's body. In some embodiments, without being bound by theory, the support structure member and / or the cells provided on the support structure help promote the growth and / or regeneration of gastrointestinal tissue of the host stem cells present at the implantation site of the support structure member.

[0014] In some aspects, the present disclosure relates to the following discovery: the growth of esophageal tissue can be promoted or enhanced by the presence of a synthetic support structure, which is designed to replace or repair the natural structural pattern and / or functional characteristics of a diseased or injured tissue or organ without the need for the support structure member to be fully integrated into the final regenerated tissue. Therefore, in some aspects, the present disclosure provides a method for promoting or enhancing the growth of gastrointestinal (e.g., esophageal) tissue, the method comprising: delivering a synthetic support structure member to a gastrointestinal (e.g., esophageal) region of a subject, wherein the delivery of the synthetic support structure causes the growth of new gastrointestinal (e.g., esophageal) tissue in the region of the subject. In some embodiments, the diseased or injured gastrointestinal region is resected (e.g., by surgery) before the support structure member is implanted. In some embodiments, the support structure is an implanted approximately tubular structure (e.g., sutured to the end of the remaining gastrointestinal tissue after removing the diseased or damaged tissue). In some embodiments, the implanted support structure is shorter than the removed tissue (e.g., 5-50% shorter). In some embodiments, when the tissue is attached (e.g., sutured) to both ends of the support structure, the remaining gastrointestinal tissue near the implant site is stretched. In some embodiments, new gastrointestinal tract (e.g., esophagus) tissue is regenerated on the implanted support structure, but is not fully integrated with the support structure. In some embodiments, even if the support structure member can be retained in the cavity of the regenerated tissue, the wall of the regenerated tissue will not contain the wall of the support structure. In some embodiments, the support structure member can be removed from the inner cavity formed by the regenerated tissue at an appropriate point in the tissue regeneration process.

[0015] In some embodiments, the growth of new gastrointestinal tract (eg, esophageal) tissue results in the formation of functional tissue (eg, a functional esophagus) that can function without the continued presence of a supporting structure.

[0016] In some embodiments, the synthetic support structure member is absorbable or dissolvable under physiological conditions.In some embodiments, after the functional esophagus is formed, the synthetic support structure member is removed from the gastrointestinal tract (eg, esophagus) region of the subject.

[0017] In some embodiments, the methods and compositions described herein can also be used for tracheal and / or bronchial tissue regeneration.

[0018] In certain embodiments, the synthetic support structure member is used in a procedure involving connecting two separate gastrointestinal tissues to one another, either in direct contact with one another or by placing the synthetic support structure member in a bridging relationship between the respective regions. In certain embodiments, the two separate gastrointestinal tissues may be the esophagus and stomach; the stomach and small intestine; the small intestine and the large intestine. In certain embodiments, the two separate regions of tubular gastrointestinal tissue may be the esophagus and stomach, as occurs in a procedure such as a gastric lift.

[0019] These and other aspects are described in greater detail herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] The present disclosure can be best understood when the following detailed description is read in conjunction with the accompanying drawings. It should be emphasized that, according to common practice, the various features in the drawings are not drawn to scale. Instead, the sizes of the various features are arbitrarily enlarged or reduced for clarity.

[0021] Figure 1A is a perspective view of an embodiment of a composite support structure member disclosed herein, a portion of which is shown in partial cross-section;

[0022] Figure 1B is a surface micrograph of a tube surface of an embodiment of a composite support structure member disclosed herein;

[0023] Figure 1C is a side perspective view of a second embodiment of a composite support structure member disclosed herein;

[0024] Figure 2 is a non-limiting description of the esophageal biolayer; and

[0025] Figure 3 Non-limiting examples of regenerated esophageal tissues are shown compared to corresponding native tissues;

[0026] Figure 4Ais a SEM micrograph of an outer surface region of an embodiment of a synthetic support structure member disclosed herein, taken at 5000X, showing cell growth after seven days of bioreaction;

[0027] Figure 4B is a photomicrograph of an outer surface region of an embodiment of a synthetic support structure member disclosed herein showing cell growth after seven days of bioreaction;

[0028] Figure 5 is a schematic flow chart of a first embodiment of the regeneration method disclosed herein;

[0029] Figure 6 is an overall research process of an embodiment of the method disclosed herein, including the generation of a cellularized support structure component and subsequent implantation;

[0030] Fig. 7A 1 is a SEM of a sample of an electrospun support structure member according to an embodiment disclosed herein, taken at 1000X, 2000X and 5000X respectively;

[0031] Figure 7B is a graphical depiction of representative uniaxial mechanical testing loads of an electrospun support structure member before and after implantation according to embodiments disclosed herein;

[0032] Figure 7C is a table of uniaxial mechanical properties of support structure members prepared according to embodiments disclosed herein before and after implantation;

[0033] Figure 8 is a schematic diagram of flow cytometry of MSCs isolated from adipose tissue and proliferated up to 5 passages;

[0034] Fig. 9 is an overview of an implantation procedure according to embodiments disclosed herein;

[0035] Fig.10 is a representation of a timeline according to an embodiment of the process disclosed herein;

[0036] Fig.11A is a photograph of regenerated esophageal tissue in the esophagus of the first test subject, the tissue being located at the esophageal resection site of the first test subject, the photograph being taken at predetermined intervals 3 to 4 weeks after the surgery after removal of an embodiment of the support structure device described herein;

[0037] Fig. 11B is located in Fig.11A A photograph of regenerated tissue inside the esophagus at the esophagectomy site shown, taken at an intermediate time point after removal of the support structure device, showing tissue growth;

[0038] Fig. 11C is located in Fig.11AThe photo shows the regenerated tissue inside the esophagus at the site of esophagectomy. Fig. 11B Subsequent time points after the intermediate time point show tissue growth;

[0039] Fig. 12A A photograph of regenerated tissue inside the esophagus of a second test subject, the tissue being located at a esophageal resection site of the second test subject, the photograph being taken 3 to 4 weeks after removal of an embodiment of a support structure device described herein;

[0040] Fig. 12B is located in Fig. 12A Photograph of regenerated tissue inside the esophagus at the esophagectomy site shown, taken at an intermediate time point after removal of the support structure, showing tissue growth;

[0041] Fig. 12C is located in Fig. 12A Photograph of regenerated tissue inside the esophagus at the esophagectomy site shown, taken at an intermediate time point after removal of the support structure, showing continued Fig. 12B Tissue growth following that shown in ;

[0042] Fig.12D is located in Fig. 12A Photograph of regenerated tissue inside the esophagus at the esophagectomy site shown, taken at an intermediate time point after removal of the support structure, showing continued Fig. 12C Tissue growth following that shown in ;

[0043] Fig.12E is located in Fig. 12A Photograph of regenerated tissue inside the esophagus at the esophagectomy site shown, taken at an intermediate time point after removal of the support structure, showing continued Fig.12D Tissue growth following that shown in ;

[0044] Fig.13A is a tissue photograph of the esophagus of a representative test animal 2.5 months after implantation, including the surgical site and adjacent distal and proximal tissues removed for histological analysis;

[0045] Fig. 13B It is taken from Fig.13A An enlarged cross-sectional photograph of a mucosal tissue sample of the proximal portion 1;

[0046] Fig. 13C It is taken from Fig.13A An enlarged cross-sectional photograph of a mucosal tissue sample of the proximal portion 2;

[0047] Fig.13D It is taken from Fig.13A A magnified cross-sectional photograph of a submucosal tissue sample of the proximal portion 1;

[0048] Fig.13E It is taken from Fig.13A A magnified cross-sectional photograph of a submucosal tissue sample of the proximal portion 2;

[0049] Fig.13F It is taken from Fig.13A An enlarged cross-sectional photograph of a mucosal tissue sample of the distal portion 3;

[0050] Figure 13G It is taken from Fig.13A an enlarged cross-sectional photograph of a mucosal tissue sample of the distal portion 4;

[0051] Fig.13H It is taken from Fig.13A an enlarged cross-sectional photograph of a submucosal tissue sample of the distal portion 4;

[0052] Fig.13I It is taken from Fig.13A an enlarged cross-sectional photograph of a submucosal tissue sample of the distal portion 4;

[0053] Fig.14A is a photograph of a histological analysis of porcine esophageal tissue 2.5 months after implantation of an embodiment of a support structure member disclosed herein;

[0054] Fig. 14B It is taken from Fig.14A A magnified cross-sectional photograph of the B portion of the sample, illustrating the presence of mucosal tissue;

[0055] Fig. 14C It is taken from Fig.14A A magnified cross-sectional photograph of the sample from Part C, illustrating the presence of mucosal tissue;

[0056] Fig.14D It is taken from Fig.14A A magnified cross-sectional photograph of a sample from part D of FIG. 1 , illustrating the presence of mucosal and submucosal tissue and muscle layer;

[0057] Fig.14E It is taken from Fig.14A A magnified cross-sectional photograph of the sample from part E of FIG. 1 , illustrating the presence of mucosal and submucosal tissue and muscle layer;

[0058] Fig.14F For Ki67 immunoreactivity analysis Fig.14A Cross-sectional photographs of esophageal tissue samples;

[0059] Figure 14G For CD31 immunoreactivity analysis Fig.14A Cross-sectional photographs of esophageal tissue samples;

[0060] Fig.14H For CD3ε immunoreactivity analysis Fig.14A Cross-sectional photographs of esophageal tissue samples;

[0061] Fig.14I For ɑSMA immunoreactivity analysis Fig.14A Cross-sectional photographs of esophageal tissue samples;

[0062] Fig.14J For Transgelin / SMA22ɑ immunoreactivity analysis Fig.14A Cross-sectional photographs of esophageal tissue samples;

[0063] Figure 14K yes Fig.14A Photographs of cross-sections of esophageal tissue samples;

[0064] Fig.15A is a photograph of the supporting structural components and stent 21 days after implantation;

[0065] Fig. 15B is an endoscopic image showing the lumen of a patent tissue fibrovascular tube after removal of the supporting structural member and stent;

[0066] Fig.16A is a representative gross image of an explanted esophagus containing a treatment site at necropsy. The esophagus of a test animal treated with an embodiment of a synthetic support structure member disclosed herein (30-day survival), arrows indicate areas that were not epithelialized, scale bar is 5 cm;

[0067] Fig. 16B is a representative gross image of an explanted esophagus containing a treatment site at necropsy. The esophagus of a test animal (90-day survival) treated with an embodiment of a synthetic support structure member disclosed herein, scale bar is 5 cm;

[0068] Fig. 16C is a representative gross image of an explanted esophagus containing a treatment site at necropsy. The esophagus of a test animal (365 days survival) treated with an embodiment of a synthetic support structure member disclosed herein, scale bar is 5 cm;

[0069] Fig.16D Representative gross images of the esophagus containing the treatment site removed at necropsy from a control animal that survived 30 days. Arrows indicate areas that were not epithelialized. Scale bar is 5 cm.

[0070] Fig.16E Representative gross images of the esophagus containing the treatment site removed at necropsy from a control animal that survived 30 days. Arrows indicate areas that were not epithelialized. Scale bar is 5 cm.

[0071] Fig.16FRepresentative gross images of the esophagus containing the treatment site removed at necropsy from a control animal that survived 365 days. The scale bar is 5 cm.

[0072] Fig.17A is the result of high-power microscopic histological analysis of implants removed from area 2 of the 365-day test subject and visualized using Masson's trichrome staining (MT);

[0073] Fig. 17B is the result of high-power microscopic histological analysis of implants removed from area 2 of the 365-day test subject and visualized using SM 2 staining;

[0074] Fig. 17C is the result of high-power microscopic histological analysis of implants removed from area 3 of the 365-day test subject and visualized using MT staining;

[0075] Fig.17D Results of high-power microscopic histological analysis of implants removed from the 365-day test subject area 3D and visualized using SM 2 staining;

[0076] Fig.18A are cross-sectional photomicrographs of new tissue growth generated by the control method and transplanted 30 days after implantation, stained with Mason's trichrome (MT) stain, cytokeratin-13, CK13; smooth muscle transcollin, SM22; and growth-associated protein-43, GAP43, as indicated;

[0077] Fig.18B is a photomicrograph of a cross section of new tissue growth produced by implanting a synthetic support structure member as described in Example III in a test subject and explanted 30 days after implantation, stained with Mason's trichrome (MT) stain, cytokeratin-13, CK13; smooth muscle transcollin, SM22; and growth associated protein-43, GAP43, respectively, as shown;

[0078] Fig.18C are cross-sectional photomicrographs of new tissue growth generated by the control method and transplanted 90 days after implantation, stained with Mason's trichrome (MT) stain, cytokeratin-13, CK13; smooth muscle transcollin, SM22; and growth-associated protein-43, GAP43, as shown;

[0079] Fig.18Dis a photomicrograph of a cross section of new tissue growth produced by implanting a synthetic support structure member as described in Example III in a test subject and explanted 90 days after implantation, stained with Mason's trichrome (MT) stain, cytokeratin-13, CK13; smooth muscle transcollin, SM22; and growth associated protein-43, GAP43, respectively, as shown;

[0080] Fig.18E are cross-sectional photomicrographs of new tissue growth generated by the control method and transplanted 365 days after implantation, stained with Mason trichrome (MT) stain, cytokeratin-13, CK13; smooth muscle transcollin, SM22; and growth-associated protein-43, GAP43, as shown;

[0081] Fig.18F are photomicrographs of cross-sections of new tissue growth produced by the method of implanting a synthetic support structure member as described in Example III in a test subject and explanted 90 days after implantation, stained with Mason's trichrome (MT) stain, cytokeratin-13, CK13; smooth muscle transcollin, SM22; and growth associated protein-43, GAP43, respectively, as shown; and

[0082] Fig.19 is a flow chart of a second embodiment of the regeneration method disclosed herein. DETAILED DESCRIPTION

[0083] Aspects of the present disclosure relate to the discovery that inserting a synthetic support structure member into a region of the esophagus of a subject can promote or enhance the regeneration of new esophageal tissue (e.g., a complete and functional esophagus) in the subject without requiring the support structure member to be fully incorporated into the regenerated tissue. Thus, in some embodiments, the present disclosure provides a method of promoting or enhancing the growth of gastrointestinal tract (e.g., esophageal) tissue, the method comprising: delivering a synthetic support structure member to a region of the gastrointestinal tract (e.g., esophagus) of a subject, wherein the delivery of the synthetic support structure member causes the growth of new gastrointestinal tract (e.g., esophageal) tissue in the region of the subject.

[0084] Other aspects of the present disclosure relate to the discovery that inserting a synthetic support structure member into a subject as part of a surgical procedure (e.g., gastric lift surgery, etc.) can reduce the formation of fibrotic scar tissue at or near the suture line or trauma area. Such methods and applications can be used in a procedure that includes removing a circumferential portion of the esophagus from a subject, wherein the circumferential portion of the esophagus to be removed is located between the cervical esophagus and the fundus, forming a distal cervical anastomosis and a proximal fundus anastomosis, wherein the distal cervical anastomosis and / or the proximal fundus anastomosis define the inner surface of the native tissue cavity, and replacing the removed circumferential portion with a synthetic support structure, the synthetic support structure having a first end, a second end opposite the first end, and an intermediate portion extending therebetween. At least a portion of the synthetic support structure is configured as a tubular member that defines an internal cavity and also has a polymer outer surface extending from the first end to the second end, and a cellularized layer adhered to at least a portion of the polymer outer surface. The first end of the synthetic support structure is maintained in direct contact with the distal cervical esophageal tissue and sutured to form a cervical synthetic support anastomotic connection. The second end of the synthetic support structure is directly contacted with the proximal gastric fundus tissue and sutured to form a synthetic support-gastric fundus anastomosis structure, wherein the first end and the second end are kept in contact for a period of time to achieve the growth of new esophageal tissue along the synthetic support structure, the new esophageal tissue growth originates from and contacts the tissue in the resected organ portion retained in the subject's body, and the new esophageal tissue growth occurs around the synthetic tubular support structure between the subject's cricopharyngeus muscle and the proximal gastric fundus anastomosis. After the new esophageal growth is completed, the synthetic structure will no longer be in contact with the esophagus, so that the new esophageal tissue growth is continuous and still in contact with the cervical distal region of the esophagus and the bottom proximal portion of the esophagus.

[0085] The tissue regenerated using the methods described herein can be any gastrointestinal tissue, such as the tissue of the esophagus, stomach, intestine, colon, rectum or other luminal gastrointestinal tissue. In some aspects, the present disclosure is based in part on the surprising discovery that the methods described herein allow the regeneration of gastrointestinal tissue comprising muscle tissue, nervous system tissue, or muscle tissue and nervous system tissue.

[0086] In some embodiments, the synthetic support structure member is absorbable or dissolvable under physiological conditions (e.g., within a time period approximately corresponding to the time required for tissue regeneration). In some embodiments, at least a portion of the synthetic support structure member is absorbable or dissolvable under suitable physiological conditions.

[0087] In some embodiments, after the regenerated functional tissue (eg, esophagus or portion thereof) is formed, the synthetic support structure member is removed from the subject.

[0088] In some embodiments, the composite support structure member is part of a system that includes a bracket or other pressure device configured to be positioned inside an inserted composite support structure member.

[0089] In some embodiments, the stent can be positioned while the synthetic support structure member is inserted. In some embodiments, the stent can be removed from the subject after the regenerative functional tissue (e.g., esophagus or portion thereof) is formed. In some embodiments, the stent can be removed while the synthetic support structure member is removed. In some embodiments, the stent can be removed or repositioned while the synthetic support structure member remains in place. In certain embodiments, the stent can be replaced with an additional stent while the synthetic support structure member remains in place. In certain embodiments, the stent can be placed after the synthetic support structure member is removed from the subject after the regenerative functional tissue (e.g., esophagus or portion thereof) is formed.

[0090] In some embodiments, the support structure member is designed to be easily retrievable by having: a) one or more reversible attachments that are easier to remove than sutures, for example to facilitate disconnection of the support structure member from surrounding tissue (e.g., esophagus) after tissue regeneration, and / or b) one or more features that can be used to assist in retrieval of the support structure member, for example after it has been disconnected from surrounding tissue (e.g., adjacent esophageal tissue).

[0091] Non-limiting examples of reversible attachments include mechanical mechanisms (e.g., hooks and loops, connectors (e.g., stents), or other mechanical attachments that can be disconnected) and / or chemical mechanisms (e.g., biodegradable or absorbable attachments and / or attachments that can be selectively removed by chemical or enzymatic methods). In some embodiments, absorbable staples can be used. In some embodiments, absorbable staples include, for example, poly(lactic-co-glycolic acid), or any other absorbable material mixture.

[0092] In some embodiments, surgical implantation and / or retrieval of support structure members may be performed with the assistance of a thoracoscope.

[0093] Non-limiting examples of structural features that can assist in retrieving or removing a support structure member (e.g., after it is disconnected from the surrounding gastrointestinal tissue) include openings, indentations, protrusions, or other structural features, or any combination thereof, that are located only on the outer surface of the support structure member. One or more of these structural features can be used to help grasp or hold a tool (e.g., a grasper) for retrieving the support structure member. In some embodiments, one or more of these structural features can be located only at one end of the support structure member (e.g., an end near the subject's mouth). In some embodiments, one or more of these structural features can be located at both ends of the support structure member, or on its entire length. In some embodiments, one or more of these structural features are located only on the outer surface of the support structure member. In some embodiments, one or more of these structural features are located only on the inner surface of the support structure member. In some embodiments, one or more of these structural features are located on the outer and inner surfaces of the support structure member. In some embodiments, the support structure member is reinforced (e.g., thicker and / or includes a stronger material) at or around the location of one or more structural features for retrieving the support structure member.

[0094] In some embodiments, the disconnected support structure member can be removed by endoscopic inspection of the airway lumen leading to the esophagus. In some embodiments, the disconnected support structure member can be removed surgically.

[0095] In some embodiments, the subject has disease (e.g., cancer) or injured gastrointestinal tissue that needs to be replaced. In some embodiments, the subject is a human (e.g., a human patient). In some embodiments, the disease can be one of esophagitis, refractory chronic stricture, damage caused by caustic burns, perforation, or other damage that may lead to end-stage organ dysfunction requiring surgical repair.

[0096] In some embodiments, the subject has a congenital anomaly, such as a congenital anomaly of the esophagus, such as esophageal atresia (EA), including but not limited to long gap esophageal atresia (LGEA).

[0097] In some embodiments, the present disclosure provides an engineered support structure member that can be used to replace or repair the esophagus or a portion thereof. In some embodiments, the esophageal support structure member described herein can be used to promote tissue regeneration (e.g., a regenerated esophagus or a portion thereof) to replace tissue in a subject (e.g., a human being). For example, a subject (e.g., a human being) suffering from certain cancers (e.g., esophageal cancer) may benefit from replacing tissue or organs affected by cancer. Without being bound by any particular theory, the synthetic support structure member described herein promotes the growth of new tissue (e.g., esophageal tissue) in a subject, thereby providing a therapeutic benefit to the subject.

[0098] In some embodiments, the growth of new esophageal tissue results in the formation of a functional esophagus and or an esophageal organ junction in a subject. In some embodiments, the new esophageal tissue does not incorporate the support structure members into the regenerated esophageal wall. In some embodiments, the support structure members are designed and manufactured to be absorbable and / or easily retrievable after esophageal tissue regeneration. In some embodiments, the support structure members are designed to be at least partially absorbable.

[0099] In some embodiments, the size and shape of the synthetic support structure member approximates the size and shape of the diseased or injured gastrointestinal tract (eg, esophagus) region being replaced.

[0100] In some embodiments, the synthetic support structure member may be used in corrective surgery where a portion of the stomach is reshaped and connected to the proximal end of the esophagus.

[0101] In some embodiments, the support structure member will have at least two layers. In certain embodiments, the support structure member may have an approximately tubular structure. Figure 1A A non-limiting embodiment of a support structure member 10 is shown, which has an approximately tubular body 12, the body 12 having an inwardly oriented surface 14 and an outwardly oriented surface 16. In some embodiments, the cross section of the support structure member 10 is approximately circular. In some embodiments, the cross section is approximately "D" shaped. However, support structure members 10 with other cross-sectional shapes can also be used. The support structure member 10 can have any suitable length and diameter, depending on the size of the corresponding tissue being regenerated. In some embodiments, the length of the support structure member 10 can be about 1-10 cm (e.g., 3-6 cm, for example, about 4 cm), or 10-20 cm long in other embodiments. However, it is conceivable that shorter or longer support structure members 10 can be used depending on the application, the needs of the patient and / or the location in the gastrointestinal tract that needs to be treated. In some embodiments, the support structure member 10 may have an inner diameter of 0.5 to 5 cm. However, depending on the application, the needs of the patient and / or the location of the gastrointestinal tract that needs to be treated, a support structure member with a smaller or larger inner diameter can be used.

[0102] In some embodiments, the length of the support structure member 10 can be shorter than the length of the gastrointestinal tract (e.g., esophagus) region being replaced or replicated. In some embodiments, the length of the support structure member 10 is 50-95% (e.g., about 50-60%, 60-70%, 70-80%, 80-90%, about 80%, about 85%, about 90%, or about 95%) of the length of the tissue being replaced. Without being bound by any theory, it is believed that certain areas of the relevant gastrointestinal tract region can actively respond to the traction forces applied to the relevant organ tissue, thereby generating certain bioorganically mediated signals, thereby initiating or promoting tissue growth and differentiation.

[0103] In certain embodiments, the length of the support structure 10 can be longer than the length of the replaced gastrointestinal tract (e.g., esophagus) region. In some embodiments, the length of the support structure 10 is between 100% and 150% of the replaced length (e.g., about 100-110%, 110-120%, 120-130%, 130-140%, about 100%, about 105%, about 110%, or about 115%). It can be considered that the length of the support structure is necessary to effectively replace the affected area. In some cases, it is envisioned that the length of the support structure 10 will be longer than the replaced gastrointestinal tract region to effectively position the support structure and reduce trauma and ischemia in the affected or associated area.

[0104] In some embodiments, support structure 10 may be composed of a single layer of synthetic material. However, within the scope of the present disclosure, support structure 10 may also include multiple layers of synthetic material.

[0105] Thus, in some embodiments, the synthetic support structure 10 can be composed of multiple layers (e.g., two or more layers, such as two, three, four, five or more layers). In some embodiments, one or more layers are made of the same material. In some embodiments, different layers are made of different materials (e.g., different polymers and / or different polymer arrangements). The synthetic support structure 10 as disclosed herein can include two or more different components that are assembled to form an existing support structure, such as before cellularization and / or implantation. In some embodiments, the synthetic support structure 10 includes two or more layers in contact with each other, such as by a synthetic technique for making the support structure 10. In some embodiments, the support structure 10 can be synthesized using a technique involving multiple steps of combining two or more layers together, such as applying a layer of electrospun material to a portion of a previously made support structure, such as a previously electrosprayed material layer, a previously electrospun material layer, a surface of a different component (e.g., a braided tube or mesh) incorporated into the support structure, or a combination of two or more thereof.

[0106] exist Figure 1AIn the illustrated embodiment, the support structure 10 includes at least one outer layer 18 that defines the outer surface 14 of the support structure body 12. The support structure 10 includes at least one additional inwardly oriented layer 20. In the illustrated embodiment, the at least one additional inwardly oriented layer 20 is in direct contact with the inwardly oriented face of the outer layer 18. When needed or desired, the at least one inwardly oriented layer 20 can be configured to provide structural support to the associated support structure body 12. Figure 1A In the illustrated embodiment, the at least one inwardly oriented layer 20 may be configured as a suitable mesh or braid that is positioned circumferentially around at least a portion of the longitudinal length of the support structure body 12. In other embodiments, it is contemplated that the at least one inwardly oriented layer 20 may be comprised of a suitable polymer layer. Figure 1A In the illustrated embodiment, the body 12 of the support structure 10 includes at least one layer 22 positioned within the mesh or woven layer 20 .

[0107] When needed or required, the support structure 10 can have a generally uniform wall thickness. However, in some embodiments, the wall thickness can vary at specific areas of the body 12. In some embodiments, the wall thickness at one or both ends 24, 26 of the body 12 of the support structure 10 is different (e.g., thicker) from the wall thickness of the central portion 28 of the support structure 10 (not shown). In some embodiments, the thicker wall area is stronger and provides greater support for the sutures connected to one or both ends 24, 26 of the support structure 10 when the support structure is connected to the surrounding gastrointestinal tissue. The thicker wall area can also include discrete structures that facilitate suturing. Non-limiting examples of such configurations include tubes, integral bodies, etc.

[0108] In some embodiments, at least the outwardly oriented surface 14 defined on the outwardly oriented layer 18 can be composed of an electrospun polymer material. In some embodiments, it is contemplated that the outwardly oriented wall 18 can be composed of an electrospun polymer material. In some embodiments, the electrospun outwardly oriented layer can be in direct contact with a suitable woven material layer 20. Fiber orientation

[0109] The electrospun fibers can be isotropic or anisotropic. In some embodiments, the fibers in different layers can have different relative orientations. In some embodiments, the fibers in different layers can have substantially the same orientation. In addition, the fiber orientation of each layer of the composite material or sandwich support structure can also be changed.

[0110] In some embodiments, a support structure with different porosities can be used. In some embodiments, one or more layers of a support structure allow substantially complete cell penetration and uniform inoculation. In some embodiments, one or more layers of a support structure can be constructed to prevent the penetration of one or more cell types, such as by densely packed fibers. Controlling fiber diameter can be used to change the support structure porosity, because porosity is proportional to fiber diameter. Alternatively, a mixture of different polymers can be electrospun together, and a polymer is preferentially dissolved to increase the support structure porosity. The properties of the fiber can be controlled to optimize the fiber diameter, fiber spacing or porosity, the morphology of each fiber, such as the porosity or aspect ratio of the fiber, and the shape is from circular to ribbon-like. In some embodiments, the mechanical properties of each fiber can be controlled or optimized, such as by changing the fiber composition and / or degradation rate.

[0111] In certain embodiments, electrospun fiber materials can provide Figure 1B In some embodiments, at least one electrospun layer in the support structure 10 can be a polymer fiber material, such as polycarbonate polyurethane, and can be prepared by dissolving the polycarbonate polyurethane in a suitable solvent (such as hexafluoroisopropanol, HFIP) and spinning and drying.

[0112] The spacing and porosity of the electrospun fiber material can be such that cells seeded on the surface of the support structure can adhere between the corresponding fibers in a suspended overlying relationship to allow the seeded cell material to form a thin sheet thereon, such as Figure 4A and Figure 4B shown. Layering of synthetic support structures

[0113] Aspects of the present disclosure relate to methods of producing synthetic support structures.In some embodiments, a tubular synthetic support structure (eg, a synthetic esophageal support structure) is produced on a mandrel, for example, by depositing material by electrospraying and / or electrospinning.

[0114] In some embodiments, one or more layers of the synthetic support structure provide structural support for the support structure, giving the support structure the required mechanical properties. In some embodiments, a braided material (e.g., a braided tube, such as a braid of nitinol, a PET braid, or a braid of other metal or non-metallic materials) can be inserted between two different layers of the support structure to provide structural support. The compressive force of the braided material (e.g., the force that the braid can exert on the next layer of material, such as an outer electrospun material layer) can be controlled by controlling the warp and weft density of the braid. In some embodiments, the braid can be coated (e.g., by dipping or other techniques) with an organic solvent to help attach it to one or more other layers of the support structure 10. In some embodiments, the length of the braid 20 does not extend to the end of the support structure body 12. In some embodiments, one or both ends of the support structure 10 are composed of two or more layers of material without a braided layer, and the central portion 28 of the support structure body 12 includes an additional braided layer.

[0115] In some embodiments, one or more layers of the synthetic support structure provide a barrier in the support structure, thereby creating a separation (e.g., a relatively impermeable separation) between the interior space (e.g., the lumen space) and the exterior space. In some embodiments, the barrier can be an electrosprayed polyurethane (PU) layer.

[0116] In some embodiments, the different layers of the support structure 10 can include one or more polymers (e.g., polyethylene terephthalate (PET), PU, ​​or a mixture of PET and PU). In some embodiments, the support structure 10 can include a nitinol braid sandwiched between an inner PU layer (e.g., electrosprayed or electrospun onto a mandrel) and an outer PU layer (e.g., electrosprayed onto the braided material).

[0117] In certain embodiments, a support structure or mandrel can be used to form support structure 10. In some embodiments, the support structure or mandrel can be coated with a material (eg, PLGA or other polymer) prior to depositing one or more layers of PU, PET, or a combination thereof.

[0118] In certain embodiments, the material in the woven or mesh layer may be composed of an absorbable polymer material. Support structure production - fiber materials

[0119] In some embodiments, one or more layers of supporting structure can be made of fibrous material. In some embodiments, supporting structure includes one or more types of fiber (for example, nanofiber). In some embodiments, supporting structure includes one or more natural fibers, one or more synthetic fibers, one or more polymers or their combinations in any. It should be understood that different materials (for example, different fibers) can be used in methods and compositions described herein. In some embodiments, the material has biocompatibility, so it can support cell growth. In some embodiments, the material is permanent, semi-permanent (for example, it can be continuously present for several years after implanting the host), or rapidly degradable (for example, it is reabsorbed in weeks or months after implanting the host).

[0120] In some embodiments, the support structure comprises or consists of an electrospun material (e.g., macrofibers or nanofibers). In some embodiments, the electrospun material comprises or consists of PET (polyethylene terephthalate, sometimes written as poly(ethylene terephthalate)). In some embodiments, the electrospun material comprises or consists of polyurethane (PU). In some embodiments, the electrospun material comprises or consists of PET and PU.

[0121] In some embodiments, the artificial support structure may include or consist of one or more of the following materials: elastic polymers (e.g., one or more polyurethanes (PU), such as polycarbonates and / or polyesters), acrylamide polymers, nylons, absorbable polysulfone polymers, and mixtures thereof. In some embodiments, the support structure may include or consist of polyethylene, polypropylene, polyvinyl chloride, polymethyl methacrylate (and other acrylic resins), polystyrene and its copolymers (including ABA-type block copolymers), polyvinylidene fluoride, polyvinylidene chloride, polyvinyl alcohol, in cross-linked and non-cross-linked forms with different degrees of hydrolysis (e.g., 87% to 99.5%). In certain embodiments, the polymer compound may also include a compound or method for increasing the hydrophilicity of the polymer. In certain embodiments, this may involve the addition of compounds such as block copolymers based on ethylene oxide and propylene oxide. If necessary or required, it may also be considered to increase the hydrophilicity of the polymer by suitable plasma treatment.

[0122] In some embodiments, the support structure may include or consist of a block copolymer. In some embodiments, addition polymers such as polyvinylidene fluoride, syndiotactic polystyrene, copolymers of vinylidene fluoride and hexafluoropropylene, polyvinyl alcohol, polyvinyl acetate, amorphous addition polymers (such as polyacrylonitrile and copolymers thereof with acrylic acid and methacrylate), polystyrene, polyvinyl chloride and its various copolymers, polymethyl methacrylate and its various copolymers, and PET (polyethylene terephthalate, sometimes written as polyethylene terephthalate)) can be solution spun or electrospun and combined with any other material disclosed herein to produce a support structure. In some embodiments, highly crystalline polymers (such as polyethylene and polypropylene) can be solution spun or combined with any other material disclosed herein to produce a support structure.

[0123] In some embodiments, one or more polymers are modified to reduce their hydrophobicity and / or increase their hydrophilicity after synthesis of the support structure but prior to cellularization and / or implantation of the support structure.

[0124] In certain embodiments, the diameter of the electrospun fiber may be less than 10 microns. In certain embodiments, the electrospun fiber. In certain embodiments, the diameter of the electrospun fiber may be between 3 microns and 10 microns. In certain embodiments, the diameter of the electrospun fiber may be between 3 microns and 5 microns.

[0125] In some embodiments, it is contemplated that the material in the braided layer may be made in whole or in part of a bioabsorbable material, such as PLGA, etc. It is also contemplated that in some configurations, the braided material may be loaded with materials and compounds that may promote and / or support tissue growth and regeneration. Non-limiting examples of such compounds and materials include one or more of the following: antibiotics, growth factors, etc. Electrospinning

[0126] In some embodiments, the support structure produced includes one or more layers (e.g., PU and / or PET) produced by electrospinning. Electrospun materials can be used in a variety of applications, including support structures for tissue engineering. Suitable electrospinning polymer methods may include methods described in the following literature: Doshi and Reneker. Electrospinning process and application of electrospun fibers. J Electrostat. 1995; 35: 151-60.; Reneker DH, Chun I. Nanometer diameter fibers of polymer produced by electrospinning. Nanotechnology. 1996; 7: 216-23; Dzenis Y. Spinning continuous fibers for nanotechnology. Science. 2004; 304: 1917-19; or Vasita and Katti. Nanofibers and their applications in tissue engineering. Int J. Nanomedicine. 2006; 1 (1): 15-30, wherein the contents related to electrospinning are incorporated herein by reference. Electrospinning is a versatile technique that can be used to produce randomly oriented or aligned fibers of essentially any chemical nature with diameters ranging from the nanoscale (e.g., about 15 nanometers) to the microscale (e.g., about 10 microns).

[0127] In some embodiments, the electrospinning and electrospraying techniques used herein involve charging a polymer solution (or melt) that is delivered through a nozzle (e.g., as a jet of polymer solution) and deposited on a target surface using a high voltage electric field. The target surface can be a static plate, a surface of a rotating drum (e.g., a mandrel), or other forms of collector surfaces that are both conductive and grounded so as to pull the charged polymer solution toward the surface.

[0128] In some embodiments, the electric field used is generally of the order of several kV, and the distance between the nozzle and the target surface is generally several centimeters or greater. The solvent of the polymer solution evaporates (at least partially evaporates) between leaving the nozzle and arriving at the target surface. This causes polymer fibers to be deposited on the surface. Typical fiber diameters range from a few nanometers to a few microns. The relative orientation of the fiber may be affected by the movement of the target surface relative to the nozzle. For example, if the target surface is the surface of a rotating mandrel, the fiber will be (at least partially) arranged on the surface in the direction of rotation. In some cases, the nozzle can scan back and forth between the two ends of the rotating mandrel.

[0129] In some embodiments, the size and density of the polymer fibers, the degree of fiber alignment, and other physical properties of the electrospun material may be affected by factors including, but not limited to, the properties of the polymer solution, the size of the nozzle, the electric field, the distance between the nozzle and the target surface, the properties of the target surface, the relative motion between the nozzle and the target surface (e.g., distance and / or velocity), and other factors that may affect solvent evaporation and polymer deposition.

[0130] Electrospinning and electrospraying processes can be used to produce interconnected polymer fiber support structures (eg, hollow synthetic support structures) on a mandrel. Support / spindle

[0131] In some embodiments, a support member (e.g., a solid or hollow support member) that can form the support structure 10 can be used to produce the support structure 10 (e.g., a support structure having two or more layers). For example, the support member can be an electrospinning collector, such as a mandrel, a tube, or a support of any other shape. It should be understood that the support member can have any size or shape. However, in some embodiments, the size and shape of the support member are designed to produce a support structure that supports an artificial tissue of the same or similar size as the gastrointestinal tissue (or part thereof) that is replaced or supplemented in the host. It should be understood that the mandrel used for electrospinning should have a conductive surface. In some embodiments, the electrospinning mandrel is made of a conductive material (e.g., including one or more metals). However, in some embodiments, the electrospinning mandrel includes a conductive coating (e.g., including one or more metals) covering a non-conductive central support.

[0132] Surprisingly, positioning a suitable braided material proximate the mandrel surface to be integrated into the resulting support structure 10 can be used as an aid to facilitate removal of the resulting support structure 10 from the mandrel. Support structure performance

[0133] It should be understood that aspects of the present disclosure can be used to enhance the physical and functional properties of any support structure, such as a support structure based on electrospinning and / or electrospraying fibers. In some embodiments, one or more support structure components can be a thin sheet, a cylinder, a thick rib, a solid block, a branching network, etc., or any combination of different dimensions. In some embodiments, the size of the complete and / or assembled support structure is similar or identical to the size of the tissue or organ replaced. In some embodiments, each component or layer of the support structure has a smaller size. For example, the thickness of the nanofiber layer can be from a few nanometers to 100 nanometers, to 1 to 1000 microns, or even a few millimeters. However, in some embodiments, the size of one or more support structure components can be about 1 millimeter to 50 centimeters. However, larger, smaller or medium-sized structures can be manufactured as described herein.

[0134] In some embodiments, the support structure is formed as a tubular structure, and cells can be inoculated to form a tubular tissue region (e.g., esophagus or other tubular regions). It should be understood that the tubular region can be a cylinder with a uniform diameter. However, in some embodiments, the tubular region can have any suitable tubular shape (e.g., including parts with different diameters along the length of the tubular region). The tubular region can also include a branch or a series of branches. In some embodiments, the tubular support structure produced has an opening at one end, two ends or multiple ends (e.g., in the case of a branch support structure). However, the tubular support structure can be closed at one end, two ends or all ends, and aspects of the present invention are not limited in this regard. It should also be understood that aspects of the present invention can be used to produce support structures of any type or organ, including hollow and solid organs, because the present invention is not limited in this regard. In some embodiments, aspects of the present invention can be used to enhance the stability of a support structure or other structure including two or more regions or fiber layers (e.g., electrospun nanofibers) that are not physically connected.

[0135] In some embodiments, the support structure is designed to have a porous surface having a diameter of about 10 nanometers to about 100 microns of holes, which can promote cellularization. In some embodiments, the average pore size of the hole is less than 50 microns, less than 40 microns, less than 30 microns, less than 20 microns or less than 10 microns (e.g., about 5 microns, about 10 microns or about 15 microns). In some embodiments, the average pore size of the hole is 20 to 40 microns. In some embodiments, the pore size is selected to prevent or reduce the immune response or other unwanted host response of the subject. The pore size can be estimated using calculation and / or experimental techniques (e.g., using porosity determination). However, it should be understood that holes of other sizes may also be included.

[0136] In some embodiments, the surface layer of the support structure is synthesized using fibers including one or more soluble particles, which can be dissolved during or after synthesis (e.g., by exposure to a solvent, an aqueous solution, such as water or a buffer) to leave holes of soluble particle size. In some embodiments, particles are contained in a polymer mixture pumped to the nozzle of an electrospinning device. Thus, particles are deposited with fibers. In some embodiments, the electrospinning program is configured to deposit thick fibers (e.g., with a few microns, about 10 microns and thicker average fiber diameter). In some embodiments, if the fiber is deposited in a dense pattern, one or more fibers will merge to form a larger macrostructure (e.g., 10-100 microns or thicker) before solidification. In some embodiments, these macrostructures can entangle two or more layers of fibers and / or parts (e.g., fibers) from two or more different components of a support structure, thereby increasing the mechanical integrity of the support structure. In some embodiments, when such macrostructures are formed (e.g., by electrospinning as described herein) at one or more stages during the synthesis of the support structure (e.g., to connect two or more layers and / or components), the surface of the macrostructure can be treated (e.g., etched or made porous using soluble particles as described herein) to provide a surface suitable for cellularization.

[0137] In some embodiments, the amount of flexible support structural material (e.g., slack) between two or more structural components (e.g., rings), between structural members of a single continuous structural component (e.g., arch members), and / or a woven support material can be used to determine the mechanical properties of the composite support structure (e.g., tensile strength, elongation, rotation, compression, range of motion, bending, resistance, compliance, degrees of freedom, elasticity, or any other mechanical property, or combinations thereof).

[0138] In certain embodiments, the support structure 10 may also include a cell sheath derived from cells seeded on the outer surface of the support structure during incubation. The cell sheath adheres to the outer surface of the support structure and is in overlapping relationship with it. It is envisioned that most of the cells present in the cell sheath will be connected to the outermost surface of the outer surface and span the pores defined therein to form a continuous or substantially continuous surface.

[0139] In certain embodiments, the cell sheath may have enough thickness to provide the structural integrity of the sheath layer. In certain embodiments, the cell sheath is composed of a large number of cells, which contact the outer surface of the support structure, enough to guide the regenerative cells to contact the sheath to produce a tissue wall covering the sheath but not integrated with it. In certain embodiments, the sheath may be composed of an inner lining with an average thickness between 1 and 100 cells. The cell thickness of certain embodiments may be between 10 and 100; between 10 and 30; between 20 and 30; between 20 and 40; between 20 and 50; between 10 and 20; between 30 and 50; between 30 and 60; between 40 and 60; between 40 and 70; between 70 and 90.

[0140] The support structure 10 with the associated cell sheath provides a removable insertable device that can be positioned at a suitable gastrointestinal resection site. The support structure 10 can be transported to the desired resection site together with the associated cell sheath in contact therewith for implantation. In certain embodiments, the support structure 10 is configured to be removable from the implantation site after the resected organ is properly regenerated. In certain embodiments, the removed support structure includes part or all of the cell sheath connected thereto.

[0141] Various embodiments of the method for regenerating a tubular organ (e.g., a gastrointestinal organ) are also disclosed herein. In certain embodiments, method 100 includes a resection step, which includes resecting a portion of a tubular organ in a subject, as shown in reference numeral 110. The resected organ may be a gastrointestinal tubular organ that is damaged or affected by disease injury, trauma, or congenital disease. In certain embodiments, non-limiting examples of suitable organs include one of the esophagus, rectum, etc. In certain embodiments, suitable organs include at least one of the esophagus, small intestine, colon, and rectum.

[0142] The resection can be achieved by any suitable surgical procedure, thereby producing a resected organ portion that remains connected to the gastrointestinal tract and remains in the subject after resection. In certain embodiments, the resection operation can produce suitable resection margins.

[0143] After the excision is completed, the synthetic support structure is implanted at the excision site, as shown by reference numeral 120. In some embodiments, the implantation may include the step of connecting the corresponding ends of the excised organ retained in the subject to the corresponding ends of the synthetic support structure, so that the synthetic support structure and the excised organ can be properly connected to each site. This can be achieved by one or more sutures, bio-organic tissue glue, etc.

[0144] In certain embodiments, the implanted synthetic support structure can be a tubular member having a polymer outer surface and a cellularized sheath covering at least a portion of the polymer outer surface. Various embodiments of the synthetic support structure have been discussed herein and can be adopted and utilized in the methods disclosed herein. In certain embodiments, the synthetic support structure includes a first end and a second end opposite the first end, a polymer outer surface between the first end and the second end, and a cellularized sheath covering at least a portion of the polymer outer surface. In certain embodiments, the implanting step can be a step of bringing at least a portion of the cellularized sheath into close contact with at least one resected edge of the resected organ portion.

[0145] In certain embodiments, the methods disclosed herein further include maintaining the synthetic support structure at the resection site for a period of time to achieve the step of growing guided tissue along the synthetic support structure, as shown by reference mark 130. In certain embodiments, the guided tissue growth originates from and contacts tissue in the portion of the resected organ retained in the subject. In certain embodiments, the guided tissue growth is adjacent to the relevant area of ​​the resected organ. In certain embodiments, the guided tissue growth exhibits differentiated tissue. In certain embodiments, the guided tissue growth is parallel to the outer surface of the cellularized sheath at its outward position. In certain embodiments, the guided tissue growth originates from and contacts tissue in the portion of the resected organ retained in the subject, and is adjacent to the relevant area of ​​the resected organ. The guided tissue growth will exhibit differentiated tissue growth and may be parallel to the outer surface of the cellularized sheath at its outward position.

[0146] After achieving guided tissue growth, the method 100 disclosed herein may include a step of removing the synthetic support structure, as indicated by reference numeral 140. In certain embodiments, the removal step is performed in a manner such that the guided tissue growth remains in contact with the resected organ portion remaining within the subject. In certain embodiments, the removal process may include endoscopically removing the synthetic support structure from within the guided tissue growth, as well as various endoscopic procedures.

[0147] In certain embodiments, the synthetic support structure may be composed in whole or in part of a bioabsorbable polymer material. In such cases, the methods disclosed herein may include the following steps: maintaining contact between the synthetic support structure and the resection edge for a period of time to achieve guided tissue growth along the synthetic support structure, such that at least a portion of the synthetic support structure is absorbed at the resection site within a period of time sufficient to achieve guided tissue growth along the synthetic support structure. In certain embodiments, when the support structure is composed entirely of a bioabsorbable material, the support structure is configured to maintain structural integrity during guided tissue growth. In certain embodiments, when the synthetic support structure is composed of a bioabsorbable material in a selected area, it is contemplated that the remainder of the support structure may be removed by an appropriate procedure after guided tissue growth is achieved.

[0148] The growth of guided tissue can be monitored by appropriate methods. In certain embodiments, tissue growth can be monitored endoscopically.

[0149] In certain embodiments, method 200 comprises the step of connecting a part of the first tubular organ existing in the system with the second tubular organ existing in the system.Method 200 comprises the step of preparing the first tubular organ, as shown in reference numeral 210, and the step of preparing the second tubular organ, as shown in reference numeral 220.The first tubular organ and the second tubular organ can be organs present in a system such as the gastrointestinal tract of an experimenter.In certain embodiments, suitable organ comprises at least two organs continuously existing in the system, for example esophagus, stomach, small intestine, colon, rectum.In certain embodiments, organ to be connected can be an organ in the gastrointestinal tract in sequence, for example esophagus to stomach, stomach to small intestine, small intestine to colon etc.

[0150] At least one of the two organs to be prepared can be a tubular organ of the gastrointestinal tract that is damaged or affected due to disease injury, trauma or congenital disease. In certain embodiments, the damaged tubular organ can be an esophagus. One or both of the preparation steps 210 and 220 can include removing a portion of the corresponding tubular organ. One or both of the preparation steps 210 and 220 can include forming a connecting portion of one or two related tubular organs. The formation process can be any process known to those skilled in the art. Non-limiting examples include stretching, molding, etc. to produce anastomotic sites.

[0151] When the preparation steps 210 and 220 are completed, the various anastomoses can be directly connected to each other or to the synthetic support structure 10, as disclosed herein with reference mark 230. In some embodiments, the corresponding ends of the first tubular organ and the second tubular organ can be directly connected as at the corresponding anastomoses, and the synthetic support structure 10 can be positioned in place after the corresponding ends are connected so that the synthetic support structure 10 is located below the formed connection point. In some embodiments, the synthetic support structure 10 can be inserted endoscopy and anchored by a suitable means. In some embodiments, a stent (e.g., stent 50) can be inserted into place after the synthetic support structure 10 is in place. The insertion of the stent 50 can be accompanied by other anchoring steps, such as suturing, etc. The stent insertion can be independent of other anchoring methods, which is also within the scope of the present disclosure.

[0152] In certain embodiments of the methods disclosed herein, the method may further include the step of transferring the cell material onto the polymer surface of the synthetic support structure and growing the cell material to form a cellularized material layer (e.g., a cellularized sheath), wherein the transferring and growing steps occur prior to the excision step.

[0153] In certain embodiments, the synthetic support structure used in the methods disclosed herein is a tubular member, the outer surface of which includes spun polymer fibers. In certain embodiments, the spun fibers can be electrospun by a suitable method (e.g., the method described in the present disclosure). In certain embodiments, the cellular sheath spans at least a portion of the electrospun fibers placed outward. The cellular sheath can be composed of cell material, including at least one of mesenchymal cells, stem cells, and pluripotent cells. The cell material can be from an autologous source or an allogeneic source of the subject.

[0154] Without being bound by any theory, it is believed that implanting a synthetic support structure such as disclosed in various ways herein, particularly a synthetic support structure implanted with an overlying cell sheath, can promote the growth, regeneration and differentiation of the subject's tissue in contact with or near the position of the implanted synthetic support structure. The growing regenerative tissue is guided by the synthetic support structure and the associated cell sheath to produce a tubular cell body, which is integrally connected to the remaining tubular organ excision end and flares outward to wrap the synthetic support structure and the associated cellular sheath. It is believed that the support structure and the associated cellular sheath can promote or stimulate the regeneration growth of the excised tissue while minimizing tissue rejection. It can also be believed that the presence of the cellular sheath can reduce or minimize the penetration of the regenerative tissue to the sheath during growth and differentiation. In certain embodiments, tissue generation is carried out from each end to the middle.

[0155] Without being bound by any theory, it can be believed that once implanted, the deposition of cell material on the surface of the synthetic support structure can provide signals to the native tissue (e.g., related cell microenvironment) near the anastomosis to achieve organized regeneration of differentiated tissue structure. It can also be believed that the cell material present in the cellular sheath can promote tissue remodeling, thereby reducing the incidence of fibrotic scar tissue at or near the surgical site after removing the implanted synthetic cell support. In certain embodiments, during the presence of the implanted synthetic support structure, the remodeling and / or reduction of fibrotic scar tissue can be confirmed. In certain embodiments, after removing the implanted synthetic support, the initial fibrotic scar tissue can still be remodeled and / or reduced. In certain embodiments, the remodeling and / or reduction of fibrotic scar tissue can be seen 90 days after surgery, 120 days after surgery, and 365 days after surgery. In certain embodiments, the reduction of fibrotic scar tissue formation caused by the presence of the synthetic support structure and / or the continuous induction remodeling of fibroid scar tissue can help reduce and / or eliminate stenosis at or near the surgical site.

[0156] In certain embodiments, the synthetic support structure may include other compounds that may stimulate the recipient's cells / tissue to deposit around and longitudinally throughout the synthetic structure. Non-limiting examples of such compounds may include VEGF, MMP2, and IL-8.

[0157] Once the regenerated tissue is in place, the synthetic support structure can be removed. In certain embodiments, the regenerated tissue structure will lack the inner epithelial layer immediately after the synthetic support structure is removed. Fig.11A , Fig. 11B and Fig. 11C As shown, after the support structure was removed, the layer had regenerated, and these images were taken immediately after the support structure was removed, 2 months after removal, and 3 months after removal.

[0158] The synthetic support structure disclosed herein can be advantageously used in a surgical procedure on the gastrointestinal tract of a subject, such as a surgical procedure performed between the cricopharyngeal notch and the suprasternal notch. The synthetic support structure disclosed herein can have a first end, a second end opposite the first end, and an intermediate portion extending between the first end and the second end. The synthetic support structure is configured as a tubular member defining an inner cavity, the synthetic support structure also having a polymer outer surface extending from the first end to the second end, and a cellularized layer adhered to at least a portion of the polymer outer surface.

[0159] The surgery may include removing a portion of the esophagus from the subject, in particular, removing a portion of the esophagus located between the esophageal region of the neck and the fundus to form a distal cervical anastomosis and a proximal fundus anastomosis, so that the distal cervical anastomosis and / or the proximal fundus anastomosis form a native tissue luminal surface. When needed or required, the surgery may include removing a circumferential portion of the esophagus in a defined area to form a distal cervical anastomosis and a proximal fundus anastomosis.

[0160] During the surgical procedure, a portion of the synthetic support structure disclosed herein replaces the resected portion of the esophagus, and a first end of the synthetic support structure remains in direct contact with and connected to distal cervical esophageal tissue, thereby forming a connection such as a cervical tissue-synthetic support anastomotic connection. A second end of the synthetic support structure remains in direct contact with and connected to proximal gastric fundus tissue, thereby creating a synthetic support-gastric fundus anastomosis. When necessary or desired, one or both connections can be completed by suturing.

[0161] A first end of a synthetic support structure and a second end of the support structure. The first end and the second end of the support structure are maintained in contact with the associated tissue for a period of time to achieve growth of new esophageal tissue along the synthetic support structure. The term "contact" used can be a direct end-to-end connection between one or both ends, or in certain applications can include a respective end positioned proximal to an associated anastomosis. The completed new esophageal tissue growth originates from and contacts tissue in the resected organ portion retained in the subject's body, and can occur at a location associated with a suitable stem cell niche around the synthetic tubular support structure, such as between the cricopharyngeus muscle and the proximal anastomosis of the gastric fundus of the subject.

[0162] After the new esophageal tissue growth is completed, the synthetic support structure can be removed from contact with the esophagus in a certain manner so that the new esophageal tissue growth is continuous and remains in contact with the distal portion of the neck of the esophagus and the proximal portion of the fundus of the esophagus. In certain embodiments, the synthetic support structure can be removed by using endoscopic techniques. The growth of new esophageal tissue can be guided by tissue growth such as epithelial tissue, smooth muscle tissue, vascular tissue, and neuronal cell proteins. In certain embodiments, the guided tissue growth includes epithelial tissue, smooth muscle tissue, vascular tissue, and neuronal cell proteins, which are covered on the outer surface of the tubular synthetic support structure and can be covered in a manner that does not adhere to the polymer outer surface of the tubular synthetic support structure.

[0163] When needed or required, the synthetic support structure can be part of an assembly that includes at least one pressure member, and the method can include the following steps: after surgical placement of the synthetic support structure, positioning at least one pressure member in the lumen of the synthetic support structure. Non-limiting examples of suitable pressure members include stents, nasogastric tubes, etc. The pressure member can be configured so that the end of the pressure member extends beyond one or more anastomotic sites so that after surgical placement of the synthetic support structure, the support structure is between the pressure member and the luminal surface of the native tissue. In other embodiments, when the first pressure member is in place, it spans at least one connection between at least one anastomosis and at least one end of the synthetic support structure. In certain embodiments, when the first pressure member is in place, it spans at least one connection between the corresponding anastomosis and the two ends of the synthetic support structure. In certain embodiments, the first pressure member spans the connection between the distal cervical anastomosis and the first end of the tubular synthetic support structure, and the connection between the proximal anastomosis of the fundus and the second end of the tubular synthetic support structure.

[0164] In some procedures, the synthetic support structure assembly may include a second pressure member in addition to the first pressure member discussed above. In some procedures, after guided tissue growth is completed, particularly between the cricopharyngeal notch and the suprasternal notch or other stem cell microenvironment, the first pressure member and the synthetic support structure may be removed. After removing the first pressure member, the second pressure member may be positioned at the desired location and may remain in place for a period of time to allow epithelialization of the luminal surface of the new tissue. When needed or required, the second pressure member may be configured as a stent or nasogastric tube. In some embodiments, the second pressure member is positioned in the area defined by the guided tissue growth in the esophagus at a time after the step of removing the first pressure member, and the second pressure member remains in place for at least 15 days.

[0165] In certain embodiments, the process may involve the step of transferring the proximal region of the stomach fundus of the subject to a position above the diaphragm of the subject. Examples of such processes include processes associated with abdominal pull-up surgery.

[0166] To further understand the present disclosure, reference is made to the following examples. These examples are listed for illustrative purposes and should be considered illustrative of the present disclosure and the invention set forth in the claims. Example Example 1: Esophageal support structure

[0167] like Figure 1AAs shown, the synthetic esophageal support structure comprises three layers of material. The first layer of polyurethane (PU) is deposited on a metal mandrel by electrospraying. The braided material is then deposited on the first PU layer. The second PU layer is then deposited by electrospinning. The resulting support structure is then removed from the mandrel. Each support structure defines a tubular structure having a wall comprising three layers (an inner electrospray coating, an outer electrospun layer, and a braided layer sandwiched therebetween). The physical dimensions of the support structure are determined by scanning electron microscopy (SEM). The average wall thickness of the support structure is approximately 500 microns. Figure 1B A non-limiting SEM view of a cross section of the wall is shown. Figure 1C A non-limiting visual image of a cross section of a tubular support structure is shown. The image shows that the cross section is generally "D" shaped. This can be achieved by using a mandrel with a "D" shaped cross section.

[0168] The outer electrospun layer is a layer of polymer fibers defining pores. The outer layer has an average fiber diameter of about 3 to 6 microns, an average pore size of about 15 to 20 microns, and a median pore size of about 25 to 45 microns.

[0169] The support structure is attached to a rack that is capable of rotating in a bath of liquid medium within the bioreactor chamber. The rotation mechanism may include a magnetic drive that allows the rack, with the attached support structure, to rotate about its longitudinal axis within the bath of liquid medium.

[0170] By depositing the cell solution on the outer surface of the support structure, cells (such as MSC or other stem cells) are inoculated on the support structure. The inoculated support structure is then incubated in a liquid culture medium that supports cell growth, and the method is to rotate the support structure in the liquid culture medium in the bioreactor chamber for about one week. The resulting support structure includes a cell sheath that is in a covering relationship with the outer surface of the support structure. In certain embodiments, the cell sheath may have a thickness that is enough to provide structural integrity for the sheath layer. In certain embodiments, the cell sheath is composed of a large number of cells, and these cells contact the outer surface of the support structure, which is enough to guide the regenerative cells to contact the sheath to produce a tissue wall that covers the sheath but is not integrated with it. In certain embodiments, the sheath can be composed of an inner lining with an average thickness between 1 and 100 cells. The cell thickness of certain embodiments may be between 10 and 100; between 10 and 30; between 20 and 30; between 20 and 40; between 20 and 50; between 10 and 20; between 30 and 50; between 30 and 60; between 40 and 60; between 40 and 70; between 70 and 90.

[0171] The support structure 10 with the seeded cell sheath can be implanted into the resection site and positioned in place. It is conceivable that one or more seeded cell populations present in the cell sheath can continue to grow after implantation. In this case, the seeded cells present in the cell sheath will maintain and support a structure separate from and in series with the tissue regenerated at the implantation site.

[0172] In this case, the corresponding support structure was implanted with the associated cell sheath into the esophagus of a pig. A section of the esophagus, approximately 5 cm long, was removed and replaced with a support structure section that was sutured to the end of the subject's remaining esophageal tissue.

[0173] The regeneration of the esophageal tissue was monitored endoscopy for several weeks.

[0174] The esophagus is a long muscular tube that divides the neck, chest, and abdomen. Figure 2 A cross section of the human esophagus is shown. The esophagus of an adult can be up to 18 cm to 25 cm long. The esophageal wall is made up of striated muscle at the top, smooth muscle at the bottom, and a mixture of the two at the middle. Therefore, in some embodiments, a multilayer synthetic support structure is provided herein that can promote the repair and regeneration of esophageal tissue having two or more layers corresponding to native esophageal tissue layers.

[0175] Figure 3 Shown is a stained cross section of native and regenerated esophageal tissue 1-2 weeks after implantation of an esophageal support structure in a pig. The cross section shows regeneration of essentially all esophageal tissue layers, including different muscle and glandular layers. Further analysis of the regenerated tissue showed that the support structure itself was not integrated into the regenerated esophageal wall. The support structure still exists within the esophagus, but it seems to play a guiding role in stimulating esophageal regeneration rather than becoming an integral part of the regenerated esophagus. Example 2: Esophageal Implant

[0176] like Figure 1AAs shown, the synthetic esophageal support structure produced comprises three layers, wherein the outer electrospun layer polycarbonate-polyurethane is deposited in the form of a polycarbonate polyurethane solution dissolved in hexafluoroisopropanol (HFIP) (DuPont, Wilmington, Delaware, USA) (12% w / v concentration). The electrospinning equipment used can be purchased from IME Technologies, Gaildrop, the Netherlands. The electrospun fibers are collected on a target aluminum mandrel rotating at 800 rpm and placed at a distance of 22 mm from the tip of the syringe to deposit isotropic fibers, thereby producing a support structure with an average wall thickness of 500 microns. The support structure is dried in a vacuum to remove residual solvent. The support structure is then plasma treated using a low pressure plasma system (Diener Tetra 150-LF-PC-D) through 2 consecutive cycles of ethylene and oxygen. The support structure is sterilized by gamma ray (STERIS, Northborough, Massachusetts). The dose range applied is 25-35 KGy.

[0177] The resulting tube was a polymer support structure composed of electrospun polyurethane with an outer diameter (OD) of 22 mm and a length of 11 cm.

[0178] The morphology of the electrospun fibers was analyzed by scanning electron microscopy (Zeiss-EVO MA10). The support structure samples were sputter coated using a sputter coater (Cressington-208HR, TED PELLA, Inc, Redding, CA) at 8x10 -2 Platinum and palladium were sputtered for two minutes at a pressure of 100 mbar and a potential of 300 V. The porosity was calculated using a gravimetric method. The porosity ε is defined based on the apparent density ρAPP of the fiber mat and the bulk density ρPU of the polymer: ε=l-ρAPP / ρPU. The apparent density ρAPP of the support structure is measured as a mass to volume ratio on a 10 mm dry disk: ρAPP=mass / VPU. The pore size was measured using a mercury porosimeter system (Micromeritics AutoPore IV). Tensile tests were performed on 10 mm x 40 mm samples mounted on an electromechanical load frame (Instron 5943 device) using a 1 kN load cell in accordance with ASTM D638 guidelines. The test parameters were the same for all samples, with a data acquisition rate of 100 Hz, a gauge length of 30 mm, and a test speed of 1 mm / sec. As Fig. 7A Scanning electron microscopy, shown at increasing magnifications, demonstrates the isotropic fiber alignment of the electrospun synthetic support structure. The smooth surface and isotropic nature of the fibers ensure that the strength and elasticity of the support structure are uniform in all directions.

[0179] Uniaxial mechanical load tensile tests were performed on three pre-implantation and three post-implantation support structures ( Figure 7B), all of which showed similar results at in vivo load values. The consistency of the six samples under in vivo loading indicates that the support structure exhibits a low degree of variability after manufacturing and in vivo implantation ( Figure 7B , Figure 7C ). Among the six support structures, the mean (±SD) tensile strain ranged between 119.5±1.61mm and 124.5±3.44mm. The tensile strain at failure of the pre-implantation sample reached 397.38%±5.52%, and the tensile strain of the post-implantation sample reached 408.61%±17.64%. Strain values ​​above 400% indicate the reliability of the manufacturing process and relative in vivo stability. The failure tensile stresses of the support structures before and after implantation were 7.25±0.59MPa and 4.43±0.77MPa, respectively. Therefore, although the elasticity of the two groups under in vivo strain was comparable, the Young's modulus of the pre-implantation sample was greater than that of the post-implantation sample ( Figure 7B , Figure 7C ). The load at failure follows the same trend as Young's modulus, with the value before implantation being greater than that after implantation.

[0180] Autologous porcine adipose-derived mesenchymal stem cells (aMSCs) were isolated and characterized from eight pigs by open fat biopsy. Eight Yucatan miniature pigs received general anesthesia and chlorhexidine skin preparation, and open adipose tissue biopsies were aseptically removed from the ventral flank. A 5-cm incision was made adjacent to the linea alba, and electrocautery was used to stop bleeding. Approximately 30–50 g of adipose tissue was isolated and transferred to a 50-mL conical tube containing α-minimum essential medium (MEM) / glutamine (Thermo Fisher Scientific, Waltham, MA) and 1% penicillin / streptomycin (Thermo Fisher Scientific).

[0181] 20-60 g of abdominal adipose tissue was surgically removed from each anesthetized Yucatan miniature pig (50-60 kg). Tissue samples were washed three times in α-minimum essential medium (MEM) / glutamine (Thermo Fisher Scientific) and 1% penicillin / streptomycin (Thermo Fisher Scientific). The washed tissue was trimmed to remove lymph nodes and blood vessels and minced into fragments less than 5 mm. Tissue fragments were separated in digestion buffer (300 IU / mL collagenase type II, 0.1% bovine serum albumin (7.5%, fraction V), 1% penicillin / streptomycin, α-MEM / glutamine) at 37°C, 5% CO2, and time for 55 minutes. After quenching in complete growth medium (StemXVivo, R&D Systems, Minneapolis, MN) and 1% penicillin / streptomycin, the cells were centrifuged at 1500 rpm for 15 minutes. The cell pellet was resuspended in 5 mL of growth medium and filtered through a 70 μm filter. The cell filtrate was centrifuged at 1500 rpm for 5 minutes. The cell pellet was suspended in 5 mL of growth medium and cells were inoculated into a culture dish according to tissue weight (3 g of adipose tissue isolate was present in each T75 flask containing 20 mL of growth medium).

[0182] Cells were washed twice with PBS without calcium or magnesium (Thermo Fisher Scientific) and then detached using TrypLe (Thermo Fisher Scientific). Cells were quenched with growth medium and centrifuged at 1000 rpm for 5 min. Cell pellets were resuspended in 1% bovine serum albumin diluted in PBS. Aliquots of 1 million cells were incubated with antibodies for 30 min at 4°C in the dark (see Supplementary Table 1 for details). Labeled cells were washed three times in buffer and secondary antibodies (Life Technologies, Carlsbad, CA) were applied as needed for 30 min at 4°C in the dark. After washing three more times, cell suspensions were placed in 96-well plates for flow cytometric analysis (Guava easyCyte HT, EMDMillipore, Billerica, MA). Live cell gating (based on viability detected by ViCount) was set based on forward scattered light (FSC) and side scattered light (SSC) values. Cell type analysis was performed using fluorescent signals compensated for unstained and isotype control antibody stained samples. The acquired data were exported and analyzed using stand-alone software (FlowJo version 10, FlowJo, LLC, Ashland, OR).

[0183] In order to evaluate colony formation, adipose-derived cells were separated and single cell suspensions were prepared according to the above description and diluted to 10 cells / mL growth medium. 100 μL cell suspensions were added to each well of a 96-well plate (Corning, Inc., New York) and the number of cells was visually inspected on the second day. After 5-7 days, visible cell colonies were formed, and the culture medium was replaced every 3 days until the colony contained at least 50 cells. The colonies present in the wells were counted and expressed as a percentage of the total wells analyzed.

[0184] The multipotency of the adipocyte separated by the ability assessment of adipogenesis and osteogenesis by chemical induction.Cells are seeded in 6-well tissue culture plates, cultured in complete growth medium, and grown to 60% or 100% confluence respectively, to carry out adipogenesis and osteogenic differentiation.After reaching confluence, culture medium is changed into adipogenic culture medium or osteogenic differentiation culture medium (CCM007, R&D Systems, Minneapolis, Minnesota).Change culture medium once every 2 days, until culture 14 days.Cells cultured in adipogenic differentiation culture medium are dyed with oil red O (American MasterTech, Lodi, California) to mark lipids, and cells cultured in osteogenic culture medium are dyed with alizarin red (EMD Millipore) to detect calcium deposition.

[0185] Glucose and lactate concentrations in bioreactor conditioned media were measured at the time of inoculation and on days 2, 5, and 7 post-inoculation (iSTAT, Abbott, Princeton, NJ).

[0186] The production of porcine cytokines and growth factors in cell supernatants was analyzed by multiplex analysis on the Luminex 200 platform or by ELISA analysis of the University of Minnesota Cytokine Reference Laboratory using commercially available kits and according to the manufacturer's instructions. The levels of porcine VEGF, GM-CSF, IL-1RA, IL-6, and IL-8 were determined using a 13-fold porcine-specific bead panel (EMD Millipore). Standard curves were interpolated from each plate using BioPlex software (BioRad, Hercules, CA) for the Luminex platform or Microplate Manager software for ELISA plates read on a BioRad 550 plate reader. All samples were measured in duplicate.

[0187] The cells were rinsed in PBS and fixed with 10% formalin for 15 minutes at room temperature. The cells were gently rinsed three times in PBS containing 0.1% Triton X-100 (PBS-T) and then incubated in 10% normal goat serum (Vector) diluted in PBS-T for 1 hour at room temperature. Rabbit anti-nestin antibody (Biolegend, 1:100) was diluted in 10% normal goat serum and PBS-T and incubated overnight at 4°C. The cells were rinsed twice in PBS-T and then incubated in fluorescent goat anti-rabbit antibody (Alexa Fluor 594, Thermo Fisher Scientific) for 1 hour at room temperature. The cells were rinsed twice and counterstained with 4′,6-diamidino-2-phenylindole (DAPI).

[0188] After 48 hours at 37°C, cells were washed twice with phosphate buffered saline (Thermo Fisher Scientific) containing calcium and magnesium and replaced with fresh growth medium. Thereafter, the medium was changed every 2 days until the confluence of the culture flask reached 70%-80%. At the time of passage, cells were separated (TrypLe, Thermo Fisher Scientific), counted (Countess, Thermo Fisher Scientific) and re-seeded at a density of 200,000 cells per T175 flask. Cells were passaged twice before inoculation of the support structure.

[0189] Each 11 cm long support structure was placed in a bioreactor and seeded with 32 million cells (viability >70%, trypan blue dye exclusion, Countess, Thermo Fisher Scientific) in growth medium supplemented with 0.1875% sodium bicarbonate (Thermo Fisher Scientific), MEM Eagle (Lonza), and 1.19 mg / mL bovine collagen (Organogenesis) dissolved in 0.01 M hydrochloric acid. The cells were incubated at 37°C, 5% CO2 for 5 minutes, and then 200 mL of growth medium was slowly added to the bioreactor. The bioreactor was incubated for 7-8 days before the support structure was implanted. The medium was changed every two days and used for various tests described below.

[0190] Porcine aMSCs were seeded onto previously characterized support structures and then incubated in a bioreactor. The seeded support structures were then implanted in Yucatan miniature pigs following esophagectomy until the support structures were removed 3 weeks later ( Figure 6), using anti-porcine CD44, CD73, CD90, CD105 and CD146 antibodies, they reproducibly stained positive for known MSC markers, while CD14, CD45, CD106, CD271 and SLA Class IIDR were negative. More than 95% of the cultured cells stained positive for nestin and aSMA, indicating that stem cell properties were maintained in culture. Multipotency was determined by chemically inducing porcine MSC isolates for adipogenesis and osteogenesis, respectively. These aMSCs were routinely expanded and characterized from passages 1 to 5 and showed consistent phenotypic and functional characteristics.

[0191] Passage 2 porcine aMSCs were seeded onto polymeric supports and incubated in a bioreactor at 37°C for 7 days (+ / - 1 day). A number of cytokines and growth factors were measured using enzyme-linked immunosorbent assays (ELISAs) to determine whether seeded aMSCs cultured on the supports secrete vascular endothelial growth factor (VEGF), granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-6, IL-8, and IL-1RA secreted by the cells were detected in the conditioned medium at levels significantly higher than those in the medium alone ( Figure 4A ). However, other cytokines, TNF-α, IL-1α, IL-1β, INF-γ, IL-10, IL-12, IL-18, platelet-derived growth factor (PDGF), and activation-regulated normal T-cell expressed and secreted (RANTES), were measured but not detected.

[0192] Section biopsies of seeded grafts were taken at the end of the 7-day incubation period to assess cell health and permeability to the support structure. Cell health was assessed by immunofluorescence staining using calcein (live cells) and ethidium bromide (dead cells). Cell identification was performed using ethidium bromide to assess cell permeability to the support structure. The presence of calcein staining in the biopsy samples indicated a population of live cells attached to the support structure. In cross-sections of the support structure biopsies, the majority of cell attachment was located on the surface of the support structure. Although there was evidence of cell proliferation and ingrowth within the support structure. Metabolic activity of the implants during bioreactor incubation was measured every 48 hours for glucose uptake and lactate production. Measurements of conditioned media consistently showed a decrease in glucose levels and an increase in lactate levels over time, both indicators of continued metabolic cell growth. In addition, cell expansion in the bioreactor over the 7-day period was quantified by total DNA content, which increased several-fold during the bioreactor cell seeding process. After 7 days of incubation, the phenotype of the cells on the support structures was further characterized, and the results showed that the cells continued to express alpha smooth muscle actin (aSMA) and nestin.

[0193] After endotracheal intubation and induction of general anesthesia, the animals were placed in the left lateral decubitus position. Hair was cut and skin preparation was performed using chlorhexidine or povidone-iodine, and the animals were covered aseptically. Standard right thoracotomy was performed at the level of the 4th intercostal space for each animal and the thoracic cavity was entered. Single-lung ventilation was achieved by using a double-lumen endotracheal tube. A 4-4.5 cm long esophagus located in the middle of the chest (behind the right hilum) was moved circumferentially and removed to form a 6 cm defect (tissue retraction at the proximal and distal ends). The inoculated support structure (6 cm long) was then implanted into the proximal and distal ends of the esophagus using polydioxanone (PDS, Ethicon Inc., Somerville, NJ) absorbable sutures. After implantation, a commercially available esophageal stent (WallFlex M00516740, Boston Scientific) was inserted under direct endoscopic guidance (Storz VideoGastroscope Silver Scope 9.3MM X 110CM, Tuttlingen, Germany). Stent deployment was performed under endoscopic and surgical visualization. The esophageal stent was fixed to the proximal and distal ends of the normal esophageal tissue using absorbable sutures.

[0194] Postoperatively, animals were placed under gastrostomy to assist feeding and maintained on a liquid diet via feeding tube for 2 weeks, a mashed diet for an additional 2 weeks, and then allowed to continue the study on solid food orally.

[0195] Approximately 21 days after implantation, the support structure was retrieved endoscopically and aMSC-impregnated platelet-rich plasma (PRP) gel was applied to improve the healing process of the newly formed esophageal conduit. After application of PRP, a new fully covered esophageal stent (WallFlex TM , 12 cm long x 23 mm outer diameter, Boston Scientific) to prevent stricture formation and maintain the anatomy during regeneration. Animals were sedated and evaluated for esophageal anastomosis and esophageal stent exchange every two weeks to directly observe the status and progress of esophageal regeneration. Follow-up observation was performed by endoscopy (Storz Video Gastroscope SilverScope 9.3MM X 110CM, Tuttlingen, Germany).

[0196] The progress of regeneration was assessed by endoscopy. After the support structure was removed, the implant area was observed endoscopically every 3-4 weeks; two representative animals are shown ( Fig.10and Figure 11). Regeneration of the mucosal layer was only partially complete 3–4 weeks after implantation. However, the process of esophageal healing continued over time, as evidenced by the formation of an initial ridge at the proximal and distal ends of the mucosal layer prior to fusion of the two layers and complete mucosal regeneration. The early reestablishment of esophageal continuity and integrity and subsequent submucosal growth from the two opposing edges of the resection were consistent in all eight animals; two animals remained in place for 8 and 9 months postoperatively and without an esophageal stent for 2 and 3 months, respectively, without signs of stenosis or stricture and with persistent oral intake and significant weight gain.

[0197] To determine the histological similarity between the regenerated and native esophageal tissue morphology, tissue samples were resected from representative porcine esophagus at 2.5 months post-implantation, including the surgical site and adjacent distal and proximal tissues, for histological analysis. Fig.13A , dotted boxes indicate specimens for histological analysis). Hematoxylin and eosin ( Fig. 13B and Fig.13D ) and Masson's trichrome staining ( Fig. 13C and Fig.13E ) Representative images of tissue sections stained with ES cells showed that the esophageal epithelium and submucosa were multilayered and intact, and the inner muscle layer had normal morphology.

[0198] Representative immunohistochemical analysis of regenerated areas is shown in Figure 14, which depicts histological analysis of porcine esophageal tissue 2.5 months after implantation of the cellularized structural supports described herein. Fig.14A A macroscopic image of the resected esophagus is depicted (with proximal suture on the left). The resected tissue samples included the surgical site, which was monitored endoscopically, and adjacent distal and proximal tissues were examined histologically (dashed boxes). Fig. 14B -E) Hematoxylin and eosin ( Fig. 14B , D) and Masson's trichrome staining ( Fig. 14C , E) Representative images of stained tissue sections. Scale bars: A = 6 cm, B, C, D and E = 200 μm. Representative immunohistochemical analysis showed Ki67 ( Fig.14F )、CD31( Figure 14G )、CD3e( Fig.14H )、aSMA( Fig.14I )、transgelin / SM22a( Fig.14J ), indicating that mucosal and submucosal cells continue to proliferate and that rhabdomyosin heavy chain (K) is relatively absent in the surgical site tissue. Scale bar: FK = 200 μm, Ki67 ( Fig.14F )、CD31( Figure 14G )、CD3e(Figure 7H), aSMA( Fig.14I )、transmyelin / SM22a( Fig.14J ), indicating that the mucosal and submucosal cells at the surgical site continue to proliferate, and rhabdomyosarcoma heavy chain ( Figure 14K The predominance of aSMA and SM22a and the relative absence of myosin heavy chain suggest that smooth muscle proliferation precedes skeletal muscle growth.

[0199] Synthetic matrices seeded with autologous mesenchymal cells (aMSCs) resulted in complete longitudinal regeneration of the resected esophagus with minimal mucosal ulceration or perforation (Table 1). All animals experienced 100% longitudinal regeneration within 2-9 weeks after graft removal, with 1 of 6 animals experiencing mucosal ulceration or perforation. No animals experienced leakage during the study. Table 1 Example 3: Other Gastrointestinal Implants

[0200] The procedures outlined in Examples 1 and 2 were carried out by replacing the region of the gastrointestinal tract located in the rectum. The results were similar to those outlined previously. Example 4: Long gap atresia repair

[0201] To simulate the repair of long gap atresia, preclinical studies in pigs were conducted in which a large portion of the native esophagus was resected and replaced with the synthetic support structure device described in Example 2 using an end-to-end anastomosis. 21 days after implantation, the support structure components that were not integrated into the developing tissue were endoscopically removed, revealing a continuous, regenerated tube of fibrovascular tissue. Endoscopic evaluation of the lumen showed that the development of the mucosal epithelial layer within the implant was fully formed within 90 days after implantation. Functionally, the animals were able to eat and gain weight. Macroscopic and microscopic histological analysis at multiple postoperative autopsy time points showed early formation of fibrovascular new tissue, followed by epithelialization of the luminal surface. Endoscopic analysis nine months after surgery also showed a complete epithelialized lumen.

[0202] A similar strategy for esophageal reconstruction was tested in a porcine model, where autologous bone marrow MSCs (BM-MSCs) were seeded onto acellular small intestinal submucosal constructs. The results of this study showed that animals that survived for more than 50 days post-implantation showed epithelialization and development of muscular structures, with the longest survival being 119 days. However, this study repaired the abdominal esophagus and used 2 surgeries, the first to allow the construct to "mature" in momentum, and then the second to repair a 3 cm full circumferential resection with the mature tissue engineered tube.

[0203] Twelve animals underwent implantation and recovered well from the surgery. Two animals in the 90-day cohort had surgery terminated prematurely. One animal was euthanized one day postoperatively due to hind limb paralysis, and the other animal was euthanized 12 days postoperatively due to pericardial effusion and lung disease. Postmortem pathology results showed that the hind limb paralysis was not caused by the surgery, and the lung disease appeared to have existed before the surgery. The other premature termination of the study was on day 297 in the 365-day trial group due to an incidental intestinal volvulus that resulted in dehydration, vomiting, and decreased body condition. The results summarized in Table 2 indicate that the use of seeded synthetic support structures is safe and feasible. Table 2

[0204] In surgical control and test animals, the initial stent was removed endoscopically on day 21 after surgery. In synthetic strut test recipients, the strut component of the synthetic strut was also removed because it adhered to the stent and released from the neotissue. (See Fig.15A ). Endoscopic visualization of the newly formed adventitial tissue revealed an intact continuous tract spanning the implantation site.

[0205] All animals were re-stented and returned to their enclosures. Animals in the 30-day group were euthanized 9 days after re-stenting and autopsy analysis showed that the tubular structure was intact and the lumen was not epithelialized in both the surgical control group and the synthetic support structure group. As indicated by the red arrow, the tissue at the implantation site was red and not epithelialized. See Fig.16A , Fig.16D as well as Fig.17A and Fig. 17B .exist 17A to 17D In the figure, E = epithelium; L = lumen; FV = fibrovascular tissue; Ad = adventitia; asterisks indicate areas of muscle cell staining in C and D. Scale bars: 1 mm in MT-stained slides (A, C), 500 μm in SM22-stained slides (B, D).

[0206] Based on the results of endoscopic examination, gross histological examination, and MT histological examination, epithelialization was fully formed in the synthetic support structure implant group and the control group at 3 months after implantation. Fig.18A and Fig.18B , Fig. 16B -F and Fig. 17C -F. In the 365-day cohort with stents, stents were removed on average at day 120. Stents in the 30-day and 90-day cohorts were in use throughout their lifespan and served to maintain the position of the synthetic construct support.

[0207] The initial stent was placed immediately after surgery and removed on day 21 after implantation. To accommodate the growth of the animals, the stent was replaced every 3–4 weeks and increased in diameter unless signs of migration or obstruction occurred. To visualize the esophageal stent, an endoscope with real-time video was used in conjunction with fluoroscopy. If the esophageal stent was removed, the esophagus was visualized after removal to determine if the esophagus was damaged in any way during the removal process and to monitor regeneration. A new stent was then inserted and deployed under fluoroscopic and endoscopic guidance. For group 1, the stent was permanently discontinued once the mucosal layer was fully formed and the stent reached a maximum diameter of 23 mm (3 to 6 months after implantation). For groups 2 and 3, the stent was used until termination. The three groups of surgical control animals (CNTL: 30 days, see Fig.18A ; At 90 days, see Fig.18C ; 365 days, see Fig.18E ) and animals implanted with the synthetic support structure described in Example 1 ( Fig.18B : 30 days; Fig.18D : 90 days; Fig.18F Esophageal tissues resected at 365 days (days 2 and 365) were further studied to assess the formation of fibrotic scar tissue. Scale bar in MT = 4 mm. Scale bar in CK13, SM22, and GAP43 = 2 mm.

[0208] exist Fig.18A -F, the upper panel stained with MT shows the resection area of ​​the surgical control group (CNTL) and the implantation area of ​​the CEI recipient animals at 30, 90, and 365 days after surgery. Fig.18B , Fig.18D and Fig.18F Tissue zones are marked in the figure, including zone 1 - native tissue on both sides of the resection area; zone 2 - transition zone; zone 3 - central fibrovascular tissue.

[0209] Cytokeratin (CK13) immunohistochemistry (IHC) shows a cytokeratin-positive epithelial layer spanning the resected portion of the synthetic support structure implant (green arrows). SM22 IHC identifies smooth muscle components in the native tissue as well as in the regenerated area of ​​the synthetic support structure implant recipient (annotated arrows). SM22 also identifies vascular structures throughout the resected area and within the synthetic support structure implant area (red arrows). GAP43 IHC identified growth cone-positive neuronal structures at the border of the resected area and at the proliferative front of the regenerated smooth muscle tissue that migrated into the synthetic support structure implant area (note the arrows within the boxes). The boxed area in the SM22 IHC panel represents the area in the GAP43 IHC panel. Fig.18E In the figure, the brackets marked with asterisks indicate the resection area of ​​the surgical control group, which showed persistent non-regenerating fibrovascular tissue throughout zone 2 at 1 year after surgery, demonstrating the presence of fibrotic scar tissue. This was not evident in the resection tissue of the synthetic support structure implantation area.

[0210] Several embodiments have been described above for various aspects of the present invention, and it should be understood that various changes, modifications and improvements will be easily conceived by those skilled in the art. Such changes, modifications and improvements are intended to be part of the present disclosure and are intended to be within the spirit and scope of the present invention. Therefore, the foregoing description and drawings are only intended to be examples.

[0211] The indefinite articles "a" and "an" as used in this specification and claims, unless expressly stated otherwise, should be understood to mean "at least one".

[0212] The phrase "and / or" used in this specification and claims should be understood to mean "either or both" of the elements so combined, that is, the elements appear in combination in some cases and in separation in other cases. In addition to the elements explicitly identified in the "and / or" clause, other elements may optionally be present, whether related or unrelated to the elements explicitly identified, unless the contrary is explicitly stated. Therefore, as a non-limiting example, when used in conjunction with open language, it may refer to the following situations: for example, "comprising" may mean including A but not B (optionally including elements other than B) in one embodiment; including B but not A (optionally including elements other than A) in another embodiment; including both A and B (optionally including other elements) in yet another embodiment; and so on.

[0213] As used in this specification and claims, "or" should be understood to have the same meaning as "and / or" defined above. For example, when separating items in a list, "or" or "and / or" should be interpreted as inclusive, that is, including at least one, but also including multiple or a group of elements, as well as optional other items that are not listed. Only terms that clearly indicate the opposite meaning, such as "only one" or "exactly one", or "consisting of..." used in the claims, refer to the inclusion of exactly one element in a plurality or a series of elements. In general, the term "or" used in this article should only be interpreted as indicating an exclusive choice (i.e., "one or the other but not two") when it is preceded by an exclusive term, such as "either", "one of them", "only one of them" or "exactly one of them". "Substantially consisting of...", when used in the claims, should have the usual meaning in the field of patent law.

[0214] As used in this specification and claims, the phrase "at least one" when referring to a list of one or more elements should be understood to refer to at least one element selected from any one or more elements in the list of elements, but does not necessarily include at least one of each element specifically listed in the list of elements, nor does it exclude any combination of elements in the list of elements. This definition also allows for the optional presence of elements other than the elements explicitly identified in the list of elements referred to by the phrase "at least one", whether related or unrelated to the explicitly identified elements. Thus, as a non-limiting example, "at least one of A and B" (or equivalently, "at least one of A or B", or equivalently, "at least one of A and / or B") may refer to at least one, optionally including more than one A, without B (and optionally including elements other than B) in one embodiment; to at least one, optionally including more than one B, without A (and optionally including elements other than A) in another embodiment; to at least one, optionally including more than one B, without A (and optionally including elements other than A) in yet another embodiment; and to at least one, optionally including more than one A, and at least one, optionally including more than one B (and optionally including other elements); and so on.

[0215] In the claims and the foregoing specification, all transitional phrases such as "comprises," "comprising," "carrying," "having," "containing," "involving," "holding," and the like shall be construed as open-ended, i.e., including but not limited to. Only the transitional phrases "consisting of" and "consisting essentially of" shall be closed or semi-closed transitional phrases, respectively, as provided in Section 2111.03 of the U.S. Patent Office Manual of Patent Examining Procedure.

[0216] The use of ordinal numbers such as "first", "second", "third", etc. in the claims to modify claim elements does not in itself indicate the priority, precedence or order of one claim element relative to another claim element, nor does it indicate the temporal order of performing method actions. It is merely used as a label to distinguish one claim element with a specific name from another element with the same name (but using an ordinal number), thereby distinguishing the claim elements.

[0217] While the present disclosure has been described in conjunction with certain embodiments, it should be understood that the present disclosure is not limited to the disclosed embodiments, but on the contrary, the present disclosure is intended to cover various modifications and equivalent arrangements within the scope of the appended claims, which scope should be given the broadest interpretation to encompass all such modifications and equivalent structures permitted by law.

Claims

1. A method for reducing the remodeling formation of fibrotic tissue in a tubular organ, the method comprising the following steps: resecting a portion of a tubular organ of a subject, thereby producing a resected organ portion, the resected organ portion remaining within the subject; removing a circumferential portion of the tubular organ from the subject, replacing the removed circumferential portion at the implantation site with a synthetic support structure having a first end, a second end opposite the first end, and an intermediate portion extending between the first end and the second end, wherein at least a portion of the synthetic support structure is configured as a tubular member defining an interior lumen, the synthetic support structure further having a polymeric outer surface extending from the first end to the second end, and a cellularized sheath adhered to at least a portion of the polymeric outer surface; maintaining the first end of the synthetic support structure in direct contact with the distal portion of the tubular organ and suturing them, thereby creating a first tubular organ-synthetic support anastomosis; maintaining the second end of the synthetic support structure in direct contact with the second portion of the tubular organ and suturing to create a first tubular organ-synthetic support anastomosis, wherein the first end and the second end are maintained in direct contact for a period of time to achieve guided tissue growth along the synthetic support structure, the guided tissue growth originating from and contacting tissue in the resected organ portion remaining in the subject, the guided tissue growth occurring around the synthetic support structure; as well as After guided tissue growth is achieved, the synthetic support structure is removed from the implantation site, the step of removing the circumferential portion of the tubular organ being performed in a manner such that the guided tissue growth is continuous and remains in contact with the resected tubular organ portion remaining within the subject.

2. The method according to claim 1, further comprising: transferring cellular material to the polymer surface of the synthetic support structure; as well as The cell material is grown to form the cellularized sheath, wherein the transferring and growing steps occur prior to the step of resecting a portion of the tubular organ.

3. The method of claim 2, wherein the synthetic support structure is a tubular member, and wherein the polymer outer surface comprises spun polymer fibers.

4. The method of claim 3, wherein the cellularized sheath spans at least a portion of the outwardly disposed electrospun fibers.

5. The method according to claim 1, wherein the cellular sheath is composed of cell materials, and the cell materials include at least one of mesenchymal cells, stem cells, and pluripotent cells. The method of claim 1 , wherein the tubular organ is a gastrointestinal organ.

7. The method of claim 6, wherein the gastrointestinal organ is the esophagus, the stomach, or the esophagus and the stomach.

8. The method of claim 1, wherein the removing step is accomplished endoscopically.

9. The method according to claim 1, comprising: resecting a portion of a tubular organ of a subject, thereby producing a resected organ portion that remains within the subject and has resection margins and forms a resection site; implanting a synthetic support structure at the resection site, the synthetic support structure having a polymer outer surface and including a first end and a second end opposite the first end, the polymer outer surface being located between the first end and the second end, a cellularized sheath covering at least a portion of the polymer outer surface, wherein at least a portion of the cellularized sheath is proximate to a resection edge of the resected organ portion; as well as maintaining contact between the synthetic support structure and the resected edge for a period of time to achieve guided tissue growth along the synthetic support structure, Wherein at least a portion of the synthetic support structure is absorbed at the resection site within a period of time sufficient to achieve guided tissue growth along the synthetic support structure.

10. The method according to claim 9, further comprising: transferring cellular material to the polymer outer surface of the synthetic support structure; as well as The cell material is grown into a cellularized layer, the transferring and growing steps occurring prior to the step of removing a portion of the tubular organ.

11. The method of claim 10, wherein the synthetic support structure is a tubular member whose outer surface comprises electrospun polymer fibers, and wherein the cellularized layer comprised of the cellular material spans at least a portion of the outwardly disposed electrospun fibers. 12 . The method according to claim 11 , wherein the cell material comprises one of a mesenchymal cell, a stem cell, and a pluripotent cell, and the cell material is derived from a subject.

13. The method of claim 9, wherein the tubular organ is a gastrointestinal organ.

14. The method of claim 13, wherein the gastrointestinal organ is the esophagus.

15. The method of claim 9, wherein the subject is a mammal.

16. The method of claim 15, wherein the mammal is a human.

17. The method of claim 10, wherein the synthetic support structure is fully absorbed.

18. The method of claim 10, further comprising monitoring tissue regeneration by endoscopy.

19. A composite support structure comprising: A body having a first end and a second end opposite the first end, the body also having at least one portion configured as a tubular member, the body including an outwardly oriented surface, the outwardly oriented surface having at least one region consisting of spun polymer fibers, the spun polymer fibers having an average fiber diameter between 15 nanometers and 10 microns, at least a portion of the spun polymer fibers being interconnected to form pores having an average pore size of less than 50 microns.

20. A synthetic support structure according to claim 19, wherein the spun polymer fibers are electrospun, interconnected and form an outer layer of the body, and the body also includes at least one inner layer, the inner layer is composed of at least one of a polymer mesh, a polymer woven support material, a solid polymer member, and an electrospun layer, and the outer layer overlaps and contacts the inner layer.

21. A synthetic support structure according to claim 20, wherein the average fiber diameter of the electrospun polymer fibers is 3 to 10 microns and is composed of at least one of the following polymer materials: polyvinylidene fluoride, syndiotactic polystyrene, copolymers of vinylidene fluoride and hexafluoropropylene, polyvinyl alcohol, polyvinyl acetate, polyacrylonitrile, copolymers of polyacrylonitrile and acrylic acid, copolymers of polyacrylonitrile and methacrylate, polystyrene, polyvinyl chloride, polyvinyl chloride copolymers, polymethyl methacrylate, polymethyl methacrylate copolymers, polyethylene terephthalate, polyurethane.

22. The synthetic support structure of claim 20, wherein at least one layer is a polymer material comprising polyethylene terephthalate, polyurethane, or a mixture of polyethylene terephthalate and polyurethane.

23. The synthetic support structure of claim 20, wherein the polymer braided support material is comprised of at least one of polyethylene terephthalate, polyurethane, nitinol, and mixtures thereof.

24. The synthetic support structure of claim 20, further comprising at least one cellularized sheath layer, the cellularized sheath layer being composed of a cellular material consisting of mesenchymal cells and stem cells present in a layer having a thickness between 1 and 100 cells.

25. The synthetic support structure of claim 24, wherein a cellularized sheath of cellular material overlies the electrospun polymer material on the polymer outer surface such that the cellular material is contained on the polymer outer surface and spans pores defined therein.

26. The synthetic support structure of claim 19 further comprises at least one opening, indentation, protrusion, or a combination thereof, defined near at least one of the first end or the second end, for assisting in retrieval of the synthetic support structure from the subject after tissue regeneration occurs around the synthetic support structure at the implantation site in the subject.

27. A method of performing a surgical procedure in the gastrointestinal tract of a subject having a cricopharyngeal notch and a suprasternal notch, the subject also having a diaphragm and a stomach having a fundus and a proximal fundus region, the method comprising the steps of: Removing a circumferential portion of the esophagus from the subject to form a resection site, wherein the esophagus has a cervical esophageal region, wherein the circumferential portion of the esophagus to be removed is located between the cervical esophageal region and the gastric fundus to form a distal cervical anastomosis and a proximal gastric fundus anastomosis, wherein the distal cervical anastomosis and / or the proximal gastric fundus anastomosis define a native tissue luminal surface; replacing the removed circumferential portion with a synthetic support structure having a first end, a second end opposite the first end, and an intermediate portion extending between the first end and the second end, wherein at least a portion of the synthetic support structure is configured as a tubular member defining a lumen, the synthetic support structure further having a polymeric outer surface extending from the first end to the second end, and a cellularized layer adhered to at least a portion of the polymeric outer surface; maintaining the first end of the synthetic support structure in direct contact with the distal cervical esophageal tissue and suturing to form a cervical-synthetic support anastomotic connection; The second end of the synthetic support structure is kept in direct contact with the proximal gastric fundus tissue and sutured to form a synthetic support-gastric fundus anastomosis, wherein the first end and the second end are kept in direct contact for a period of time to achieve the growth of new esophageal tissue along the synthetic support structure, the new esophageal tissue growth originates from and contacts the tissue in the resected organ retained in the subject, and the new esophageal tissue growth occurs around the synthetic tubular support structure between the cricopharyngeus anastomosis and the proximal gastric fundus anastomosis of the subject; as well as After achieving neo-esophageal tissue growth, the synthetic support structure is removed from contact with the esophagus in a manner such that the neo-esophageal tissue growth continues and remains in contact with the cervical distal portion of the esophagus and the fundus proximal portion of the esophagus.

28. The method according to claim 27, further comprising: transferring cellular material to the polymer outer surface of the synthetic support structure; as well as The cellular material is grown to form the cellularized layer, wherein the transferring and growing steps occur prior to replacing the removed circumferential portion with the synthetic support structure.

29. The method of claim 28, wherein the cellular material is derived from autologous stem cells harvested from a subject.

30. The method of claim 28, wherein at least a portion of the synthetic support structure comprises electrospun polymer fibers.

31. The method of claim 30, wherein the cellularized layer is composed of cell materials, and the cell materials include at least one of mesenchymal cells, stem cells, and pluripotent cells.

32. The method of claim 31, wherein the cellularized layer comprises a plurality of cells, wherein at least a portion of individual cells of the cellular material span at least a portion of the outwardly disposed electrospun fibers.

33. The method of claim 27, wherein at least a portion of the synthetic support structure is absorbed at the resection site within a period of time sufficient to effectuate guided tissue growth along the synthetic support structure.

34. The method according to claim 27, further comprising the steps of: After surgical placement of the synthetic support structure, a first pressure member is positioned within the lumen of the synthetic support structure and the luminal surface of native tissue, wherein the first pressure member is one of a stent or a nasogastric tube.

35. The method according to claim 34, further comprising the steps of: The composite support structure is removed simultaneously with the removal of the first pressure member, or the support structure is actively removed by using endoscopic techniques.

36. The method according to claim 35, further comprising the steps of: A second pressure member is positioned in the esophagus at the area defined by the guided tissue growth at a time subsequent to the step of removing the first pressure member, wherein the second pressure member is a stent or a nasogastric tube and remains in place for a period of time to allow epithelialization of the luminal surface of the new esophageal tissue.

37. The method of claim 36, wherein directed tissue growth occurs between the cricopharyngeal notch and the suprasternal notch.

38. The method according to claim 37, further comprising the steps of: The subject's gastric fundus proximal region is transferred to a position above the subject's diaphragm.

39. The method of claim 37, wherein the guided tissue growth comprises epithelial tissue, smooth muscle tissue, vascular tissue, and neuronal cell proteins, and wherein the synthetic support structure has an outer surface comprising electrospun polymer fibers.

40. The method of claim 37, wherein the guided tissue growth comprising epithelial tissue, smooth muscle tissue, vascular tissue, and neuronal cell proteins overlies the synthetic support structure but is not adhered to the polymeric outer surface of the synthetic support structure.

41. The method of claim 40, wherein the first pressure member, when in place, spans at least one connection between at least one stoma and at least one end of the composite support structure.

42. The method of claim 40, wherein when the first pressure member is in place, it spans at least one connection between a corresponding stoma and two ends of the composite support structure.

43. The method of claim 42, wherein the first pressure member spans a connection between the distal end of the cervical anastomosis and the first end of the synthetic support structure, and a connection between the proximal end of the fundus anastomosis and the second end of the synthetic support structure.

44. The method according to claim 42, further comprising the steps of: After guided tissue growth is achieved, the first pressure member is removed, wherein the first pressure member is removed before or simultaneously with the removal of the synthetic support structure.

45. The method according to claim 44, further comprising the steps of: A second pressure member is positioned in the esophagus at the area defined by the guided tissue growth at a time subsequent to the step of removing the first pressure member, the second pressure member remaining in place for at least 15 days.

46. ​​The method according to claim 45, further comprising the steps of: The proximal anastomosis of the subject's gastric fundus was transferred to a position above the subject's diaphragm.

47. The method of claim 46, further comprising: transferring cellular material to the polymer outer surface of the synthetic support structure; as well as The cellular material is grown to form the cellularized layer, wherein the transferring and growing steps occur prior to replacing the removed circumferential portion with the synthetic support structure.

48. The method of claim 47, wherein the cellular material is derived from autologous stem cells harvested from a subject.

49. The method of claim 48, wherein the cellularized layer is composed of cellular materials, and the cellular materials include at least one of mesenchymal cells, stem cells, and pluripotent cells.

50. The method of claim 49, wherein the cellularized layer comprises a plurality of cells, wherein at least a portion of the individual cells span at least a portion of the outwardly disposed electrospun fibers.

51. The method of claim 50, wherein directing tissue growth occurs between the cricopharyngeus muscle and the suprasternal notch.

52. The method of claim 44, wherein the synthetic support structure comprises at least one region comprised of electrospun polymer fibers having an average fiber diameter between 15 nanometers and 10 micrometers, wherein at least a portion of the electrospun polymer fibers are interconnected to form pores having an average pore size of less than 50 micrometers, the pores defining at least one porous region present on an outer surface of the polymer.

53. A method according to claim 52, wherein the electrospun fibers of the synthetic support structure are interconnected and form an outer layer of the synthetic support structure, and the synthetic support structure also includes at least one inner layer, the inner layer being composed of at least one of a polymer mesh, a polymer woven support material, a solid polymer member and an electrospun layer, and the outer layer is in overlapping contact with the inner layer.

54. The method of claim 53, wherein the electrospun material of the synthetic support structure has an average fiber diameter of 3 to 10 microns and is composed of at least one of the following polymer materials: polyvinylidene fluoride, syndiotactic polystyrene, copolymers of vinylidene fluoride and hexafluoropropylene, polyvinyl alcohol, polyvinyl acetate, polyacrylonitrile, copolymers of polyacrylonitrile and acrylic acid, copolymers of polyacrylonitrile and methacrylate, polystyrene, polyvinyl chloride, polyvinyl chloride copolymers, polymethyl methacrylate, polymethyl methacrylate copolymers, polyethylene terephthalate, and polyurethane.

55. The method of claim 53, wherein the electrospun material of the synthetic support structure is selected from polyethylene terephthalate, polyurethane, and a mixture of polyethylene terephthalate and polyurethane.

56. The method of claim 55, wherein the cellular material on the synthetic support structure covers and adheres to the electrospun fibers on the outer polymer surface such that the cellular material is contained on the outer polymer surface and spans pores defined therein.

57. Use of synthetic support structures in gastric lift surgery on subjects, The synthetic support structure has a first end, a second end opposite the first end, and an intermediate portion extending between the first end and the second end, wherein at least a portion of the synthetic support structure is configured as a tubular member, and wherein the synthetic support structure also has a polymer outer surface extending from the first end to the second end, and a cellular layer adhered to at least a portion of the polymer outer surface, the cellular layer being composed of a cellular material.

58. Use of a synthetic support structure according to claim 33 in gastric lift surgery, wherein the synthetic support structure comprises electrospun polymer fibers, and wherein the cellularized layer is derived from autologous stem cells harvested from a subject, and wherein the cellularized layer comprises a plurality of cells, wherein at least a portion of individual cells of the cellularized layer span at least a portion of the outwardly disposed electrospun fibers.

59. Use of a synthetic support structure according to claim 57 or 58 in a gastric lift procedure performed on a subject having a stomach with a fundus, wherein the gastric lift procedure comprises the following steps: Removing a circumferential portion of the esophagus from a mammal, wherein the circumferential portion of the esophagus to be removed is located between the esophagus and the stomach in the neck, thereby forming a distal cervical anastomosis and a proximal gastric fundus anastomosis; replacing the removed circumferential portion with the composite support structure; Maintaining the first end of the synthetic support structure in direct contact with the proximal end of the distal cervical anastomosis, and maintaining the second end of the synthetic support structure in direct contact with the proximal gastric fundus anastomosis for a period of time, so as to achieve growth of new esophageal tissue along the synthetic support structure, guide tissue growth to originate from and contact tissue in the resected organ retained in the subject, and guide tissue growth to occur around the synthetic support structure between the distal cervical anastomosis and the proximal gastric fundus anastomosis of the subject; as well as After guided tissue growth is achieved, the synthetic support structure is removed from contact with the esophagus in a manner such that the guided tissue growth continues and remains in contact with the cervical distal portion of the esophagus and the fundus proximal portion of the esophagus.

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