Tissue implants and methods for making and using same
a technology of tissue implants and implants, which is applied in the field of tissue repair, can solve the problems of undesirable platelet activation with bovine thrombin, inherent risk of transmitting unwanted agents, for example prions, and achieve the effect of increasing the temperature of the sampl
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example 1
[0055]This Example demonstrates that calcium sulfate may be used to activate platelets in platelet-rich plasma.
(a) Materials
[0056]In this Example, and in the following Examples, exothermic calcium sulfate was obtained from LIFECORE Biomedical, Chaska Minn., USA under the tradename CAPSET®, a medical grade calcium sulfate alpha-hemihydrate powder. Platelet-rich plasma was purchased from the American Red Cross, Buffalo N.Y. Bovine thrombin and Collagen Type I (from rat tail) were obtained from Sigma (Saint Louis, Mo.). Human recombinant Platelet-derived Growth Factor (rePDGF-BB) was obtained from Oncogene Research Products (Cambridge, Mass.). Anti-human Transforming Growth Factor-β1 antibody (IgG fraction of antiserum, developed in Mouse) was obtained from Oncogene Research Products (Cambridge, Mass.). Anti-human Platelet-derived Growth Factor antibody (IgG fraction of antiserum, developed in Goat), Purified Mouse IgG and Purified Goat IgG were obtained from Sigma (Saint Louis, Mo.). ...
example 2
[0076]This example again demonstrates that platelet-rich plasma can be activated by exposure to calcium sulfate. Furthermore, this example demonstrates that part of the benefit of including platelet-rich plasma in the calcium sulfate matrices can be derived in part by the release of PDGF and / or TGF-β. Samples were created as summarized in Table 2 using the procedures essentially as described in Example 1, to generate the same disk sizes as in Example 1.
TABLE 2Group NameGroup Treatment (at 37° C.)CSPBS for 1 hourCS / PRPPBS for 1 hourCS / PRP-Antibody orAnti-PDGF for 30 min thenCS / AntibodyAnti-TGF-β1 for 30 minCS / PRP-IgGGoat IgG for 30 min then mouse IgG for 30 minCS / PRP-Refrig6 hour incubation at 7-8° C.CS / PDGF-BBPBS for 1 hour
[0077]In this example, the CS / PRP group was also refrigerator-treated (CS / PRP-Refrig), because calcium sulfate precipitation reaction is an exothermic reaction and achieves, during the 30-50 minutes setting time, a temperature of 40-45° C. (Phillips, 1991, supra, ...
example 3
[0081]This example provides comparative data, which indicates that calcium sulfate-platelet-rich plasma matrices can support a similar levels of osteoblastic cell proliferation as commercially available Grafton® Putty.
[0082]Samples were prepared in accordance with the teachings of Examples 1 and 2. Grafton® Putty, a demineralized bone matrix, was obtained from Osteotech Inc. (Eatontown, N.J.). The Grafton® Putty samples (GraftonPutty) were prepared using the same polyvinylsiloxane templates: the special malleability of this bone matrix compound permitted the preparation of samples of the same standard size disk as in all other preparations.
[0083]The osteoblastic cell proliferation assay, and the data resulting therefrom was analyzed as discussed in Example 1. The results are summarized in FIG. 4. wherein human primary osteoblastic cell proliferation is indicated counts per minute (cpm). The results in FIG. 4 demonstrate that human primary osteoblastic cells cultured on CS / PRP sample...
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