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Pharmaceutical composition including laminin fragments to treat or prevent the disease, disorder or symptom of tooth dentin and/or dental pulp

a technology of laminin and composition, which is applied in the direction of skeletal/connective tissue cells, peptide/protein ingredients, inorganic phosphorous active ingredients, etc., can solve the problems of restrictive application of conventional drugs into dental treatment, dentin formation, and dentin formation, so as to promote the cell adhesion of retina pigment epithelium and/or nerves, and promote dentin formation

Pending Publication Date: 2021-07-01
TAKASHI SAITO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent text describes the problem of promoting dentin formation and the development of a drug to promote dentin growth. Conventionally, drugs that promote dentin formation do not act directly on odontoblasts, which are the cells responsible for dentin formation. The use of a laminin fragment, which promotes cell adhesion, has been explored for use in dental treatment. The invention offers a treatment or medicine containing the integrin binding fragment of laminin, either alone or with other substances like porous hydroxyapatite or type I collagen. The invention also includes a method of culturing odontoblasts in the presence of the integrin binding fragment of laminin and inducing calcification to produce hard tissue transplant materials for dental treatment. The invention provides greater effectiveness and targeted promoting dentin formation for dental applications.

Problems solved by technology

As described above, a problem for the development of drug and transplant materials to promote dentin formation (regeneration) exists, but conventional drug to promote dentin formation, for example, induces odontoblast differentiation from pulpal stem cells and mesenchymal stem cell, and does not act in odontoblasts directly.
Thus, the application of the conventional drug into dental treatment was restrictive.

Method used

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  • Pharmaceutical composition including laminin fragments to treat or prevent the disease, disorder or symptom of tooth dentin and/or dental pulp
  • Pharmaceutical composition including laminin fragments to treat or prevent the disease, disorder or symptom of tooth dentin and/or dental pulp
  • Pharmaceutical composition including laminin fragments to treat or prevent the disease, disorder or symptom of tooth dentin and/or dental pulp

Examples

Experimental program
Comparison scheme
Effect test

experiment 1

Laminin 411 Fragment (Materials and Methods)

MDPC-23 Cell

[0069]MDPC-23 cell is kindly provided by Prof. Jacques E Nor from University of Michigan. The cells were grown in Dulbecco modified eagle medium (DMEM, D5796, Sigma) supplemented with 5% fetal bovine serum (FBS, 10270-106, Gibco) (maintenance media) at 37° C. in a humidified atmosphere containing 5% CO2. For the induction of differentiation and mineralization, reagents were added on the day cells reaching confluence and changed every two days. The mineralization reagents include 5% FBS, 10 mM β-glycerophosphate (191-02042, Wako), 50 μg / mL ascorbic acid (013-19641, Wako), 100 nM dexamethasone (D2915, Sigma) and DMEM.

Laminin 411 Coating

[0070]iMatrix-411 (herein referred to as LN411-E8) (iMatrix-411, Nippi, No. 892041) was diluted by PBS and coated to tissue culture treated polystyrene (TCPS, vacuum plasma treated, hydrophilic, water contact angle 38°±9° or nontissue culture treated polystyrene (12-well plate, 351143, Falcon) (non...

experiment 2

Laminin 511 Fragment (Materials and Methods)

[0089]MDPC-23 cells were cultured in the same manner as described in Experiment 1. Laminin 511 E8 fragment was purchased from Nippi company. For comparison with other ECM protein, vitronectin (Peprotech, 140-09) was also used.

[0090]Detailed information is provided in Experiment 1 with regard to protein coating, CCK-8 assay (optimal density, cell proliferation experiment), alkaline phosphatase activity, gene expression and statistical analysis. However, in Experiment 2, both LN511-E8 and vitronectin were used for cell culture. Also, in the cell proliferation assay, data were collected on day one, day two and day four. At last, tissue culture polystyrene (TCPS) culture plates were not used in Experiment 2.

Alizarin Red Staining

[0091]At the end of the culture period (day 8), the cell monolayer was fixed with 10% neutral formalin solution (060-01667, Wako) for 20 minutes and stained with 1% alizarin red solution (pH=4.1; 011-01192, Wako) in the...

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Abstract

A pharmaceutical composition contains an integrin binding fragment for Laminin which is selected from the group consisting of Laminin 511 and Laminin 411 for treating or preventing a disease, disorder, or symptom of a tooth dentin and / or pulp or pulp tissue. A method of producing a hard tissue transplantation material includes the steps of culturing odontoblast in the presence of an integrin binding fragment for the Laminin which selected from the group consisting of Laminin 511 and Laminin 411 and obtaining a culture of odontoblast and inducing calcification or mineralization of the culture of odontoblast so as to obtain the hard tissue transplantation material.

Description

TECHNOLOGY FIELD[0001]This invention relates to a pharmaceutical composition, drug or medicine including laminin fragments to treat or prevent the disease, disorder or symptom of tooth dentin and / or dental pulp. It also relates to a method for the preparation of the hard tissue transplant materials for the tooth by culturing odontoblasts in the presence of laminin fragments. Moreover, it relates to a prevention method of caries occurrence on the surface of the tooth.[0002]The sequence listing submitted in a computer readable form under the name of “P190307US01_sequence_listing.txt” is hereby incorporated by reference into the present application. The electronic copy of the sequence listing in the computer readable form, the file size of which is 7K bytes, was created on Jul. 15, 2020.BACKGROUND TECHNOLOGY[0003]The pulp which was exposed with being untreated causes serious infection by deep caries in the pulp tissue and tooth root-surrounding tissue. The infection of the pulp tissue ...

Claims

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Application Information

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IPC IPC(8): A61K38/39A61K38/01A61K33/42
CPCA61K38/39A61K33/42A61K38/014C12N5/0654C12N2533/52A61K2300/00
Inventor TANG, JIASAITO, TAKASHI
Owner TAKASHI SAITO