A Novel Designed Chitosan Based Hydrogel Which is Capable of Consecutively Controlled Release of TGF-Beta 1 and BMP-7

Abstract

A AB BS ST TR RA AC CT T O Ob bj je ec ct ti iv ve e: : Both Bone Morphogenetic Protein 7 (BMP-7) and Transforming Growth Factor Beta 1 (TGF-β1) have stimulating effects on chondrocyte proliferation. These growth factors act at different time points of the biological repair process. We aimed to design a hydrogel, which would enable consecutive controlled release of the growth factors. M Ma at te er ri ia al l a an nd d M Me et th ho od ds s: : We designed a chitosan particle impregnated Poly (vinylalcohol) (PVA)-borax hydrogel (CPBH). CPBH allowed controlled release of BMP-7 and TGF-β1 at different time points. Hydrogel structure was analyzed by Scanning Electron Microscope (SEM). The release kinetics of Growth Factors (GFs) were determined with enzyme-linked immunosorbent assay (ELISA) and UV-spectrophotometer. Chondrocyte viability and toxicity were tested through CellTiter 96 ® Non-Radioactive Cell Proliferation (MTS) ELISA. R Re es su ul lt ts s: : The designed hydrogel showed high swelling and mucoadhesion characteristics under acidic conditions. CPBH released BMP-7 first and rapidly and TGF-β1 consecutively and slowly. It also allowed controlled release of protein/peptide based drugs for 21 days without altering their bioactive properties. At the end of 21 days, 82.62% of BMP-7 and 98.34% of TGF-β1 were consecutively released. The difference between the groups was significant (for TGF-β1, p<0.05 and for BMP-7, p<0.001). C Co on nc cl lu us si io on n: : The controlled and slow release of growth factors has been shown to be beneficial on cartilage regeneration. As it is not cytotoxic, we suggest that this hydrogel might be used in medical and pharmaceutical applications areas. K Ke ey y W Wo or rd ds s: : Delayed-action preparations; hydrogels; TGF-β1 protein, human; bone morphogenetic protein 7 Ö ÖZ ZE ET T A Am ma aç ç: : Kemik Morfojenik Proteini-7 (BMP-7) ve Transforme edici Büyüme Faktörü Beta-1 (TGF-β1)'in kıkırdak hücreleri üzerinde prolifere edici etkileri bulunmaktadır. Bu büyüme faktörlerinin etkisi biyolojik tamir sürecinin farklı zamanlarında gerçekleşmektedir. Bu çalışmada, büyüme faktörlerinin ardışık ve kontrollü olarak salınımını gerçekleştirecek bir hidrojel tasarlamayı amaçladık. G Ge er re eç ç v ve e Y Yö ön nt te em ml le er r: : Bu amaçla kitosan emdirilmiş polivinil alkol (PVA)-boraks hidrojeli (CPBH) tasarlandı. Bu hidrojel BMP-7 ve TGF-β1'in farklı zamanlarda kontrollü salınımını sağladı. Tasarladığımız hidrojelin yüzey yapısı taramalı elektron mikroskobu (SEM) ile değerlendirildi. Büyüme faktörlerinin salım kinetiği enzim aracılı immün deney (ELISA) ve ultraviyole (UV)spektrofotometre ile saptandı. Kondrosit canlılığı ve toksisite testi "CellTiter 96 ® Non-Radioactive Cell Proliferation (MTS)" ELISA yöntemiyle değerlendirildi. B Bu ul lg gu ul la ar r: : Tasarlanan hidrojel, asidik ortam koşullarında büyük oranda şişme ve mukoadezyon özellikleri sergiledi. CPBH, önce hızlı bir biçimde BMP-7'nin, sonra da yavaş olarak TGF-β1'in salınımını ardışık olarak gerçekleştirdi. İçerisindeki p...

A practical way to prepare primer human chondrocyte culture

“…However, it is not sufficient for the development of chondrocyte and other cells alone. 4,7 For this reason, FBS and ITS should also be added to RPMI-1640 or DMEM medium in order to proliferate healthy, high confluent cells in chondrocyte cultures. 7,8 FBS contains the essential extracellular matrix proteins for cellular adhesion and provides relevant hormones, enzymes, and growth factors for cellular proliferation.…”

“…PS concentration could be increased in transfer medium up to 5%, but it should be noted that antibiotics have antiproliferative effects on human tissue also. 4,7,8 ITS will stimulate cell proliferation while decreasing substantially the serum requirements for culture of many cell types. Basal media supplemented with support proliferation of chondrocyte cultures.…”

Does leukocyte-poor or leukocyte-rich platelet-rich plasma applied with biopolymers have superiority to conventional platelet-rich plasma applications on chondrocyte proliferation?

“…Thus, this biocompatible hydrogel with more stable cross-links was obtained, which behaves like an osmotic pump and maintains the controlled release of PRP. [10] Hydrogel prepared with PRP in this manner is suitable for surgical applications, and at the application area, this hydrogel lasts 21 days with no degradation. Hydrogel and chondrocyte co-culture P-PRP Group 5…”

Is there a treatment protocol in which platelet-rich plasma is effective?

“…Recent well-proved studies have reported that half-life of growth factors is maximum 2 min. [37][38][39][40] It was emphasized that when growth factors -with protein and peptide structure -in PRP obtained from human were applied after that period, bioactivity decreased and even vanished. [37][38][39][40] When the studies in which PRP was obtained from autologous blood in clinics were considered, it was found that it was prepared from 10 to 20 min.…”

“…[37][38][39][40] It was emphasized that when growth factors -with protein and peptide structure -in PRP obtained from human were applied after that period, bioactivity decreased and even vanished. [37][38][39][40] When the studies in which PRP was obtained from autologous blood in clinics were considered, it was found that it was prepared from 10 to 20 min. [7][8][9][10][11][12][13][14][15][16][17][18][19][20][21] Nevertheless, it takes time to apply it to cases, which makes its bioactive and effective treatment results controversy.…”