RhBMP-2-loaded Dopamine And Heparin Modified Hyaluronic Acid Promotes Odontogenic Differentiation Of HDPSCs

Objective:Dopamine-heparin-hyaluronic acid(DA-Hep-HA),a three-dimensional porous material,was synthesized and used as a carrier of recombinant human bone morphogenetic protein-2(rh BMP-2).To explore the effect of DA-Hep-HA loaded with rh BMP-2 on the odontogenic differentiation of human dental pulp stem cells(h DPSCs)and to provide a functional scaffold material for dental pulp regeneration.Methods:Chemically synthesis of modified material DA-Hep-HA;The grafting site and grafting efficiency of heparin and the synthesized material dopamine-hyaluronic acid were characterized by FT-IR and Elemental Analysis;Scanning electron microscopy(SEM)was used to observe the micro morphology and structure of DA-Hep-HA;Rh BMP-2 was loaded into DA-Hep-HA material by electrostatic adsorption method,and the drug loading and slow-release effect of rh BMP-2 were tested;The proliferation of h DPSCs co-cultured with the extract of DAHep-HA material was measured by MTT colorimetry to evaluate the biocompatibility of the material;Real-time quantitative PCR(q PCR)was used to detect the expression level of marker molecule genes in odontoblasts,and to evaluate the effect of rh BMP-2 dopamine-heparin-hyaluronic acid-containing material(DA-Hep-HA-rh BMP2)on the odontogenic differentiation of h DPSCs.Results:The modified material DA-Hep-HA has been synthesized successfully.It is characterized that the material has a loose and porous network structure.DAHep-HA-rh BMP2 material can slowly release rh BMP-2,and the release rate is inversely proportional to the heparin content of the material.The results of coculture of DA-Hep-HA and h DPSCs showed that it could promote the proliferation of h DPSCs,and this effect was positively correlated with the material concentration.q PCR results showed that,compared with the control group,DA-Hep-HA-rh BMP2 can up-regulate the expression of DSPP,DMP-1,Col-1 and Nestin genes.The results of q PCR showed that DA-Hep-HA-rh BMP2 could up-regulate the expression of DSPP,DMP-1,COl-1 and Nestin genes compared with the control group.Conclusion:In this study,the modified material DA-Hep-HA was loose,porous,plastic and biocompatible,which promoted the proliferation of h DPSCs.It can carry and slowly release rh BMP-2 for more than 4 weeks,and DA-Hep-HA-rh BMP-2 can promote the odontogenic differentiation of h DPSCs.DA-Hep-HA material not only provides a suitable microenvironment for stem cells such as h DPSCs,but also can be used as a carrier system of growth factors.As a functional scaffold for dental pulp regeneration,it has a potential application prospect.