Introduction: Tissue engineering is an emerging field combining material science and cell biology to modulate tissue regeneration. It is believed that native tissue microenvironment(TME) plays an important role in the process of tissue formation. Hence, a bioscaffold mimicking the niches of native TME can definitely accelerate tissue regeneration. In our previous study, we successfully replicated the surface topography, mechanical and biochemical properties of tendon tissue section into Polydimethylsiloxane(PDMS) material through a technique named of bio-imprinting. The study proposed that the mesenchymal stem cells(MSCs) could effectively and precisely differentiate into tenocytes, when the scaffold was specifically designed with correct biochemical information, accurate surface topography and appropriate mechanical property of tendon TME. In the current study, we aim at translating these significant findings into an implantable membrane called click-chemistry based polymer poly1 ,8-octanediol citrate(POC-click). for tendon regeneration. A series of characterizations has been used to evaluate the feasibility of this novel and implantable membrane. Materials and Methods: Membrane made of POC-click was fabricated through the established technique in which it was able to replicate the surface pattern and mechanical property of tissue. Type Ⅰ Collagen was coated onto the membrane to serve as biochemical component of TME. Hydrophilicity of membrane was tested through water contact angle measurement. Surface topography was characterized by SEM and AFM.respectively.To investigate cellular behaviors, mesenchymal stem cells(MSCs) were cultured on the polymer membrane and natural tendon section, respectively. H&E staining was applied in order to compare the morphology and alignment of MSCs. Tenomodulin antibody was applied in the immunohistological chemical(IHC) staining so as to verify the tenogenic differentiation of MSCs. Results and Discussion: Water contact angle measurement (Figure 1 A) illustrates that membrane made of POC-click exhibits significantly lower contact angle as compared with PDMS scaffold, indicating that the POC-click membrane possesses superior hydrophilicity and cell affinity than the PDMS material. The SEM image(Figure 1B) demonstrates that the topographical information of closely packed collagen fibrils in bovine tendon section was successfully replicated into the POC-click membrane by bio-imprinting technique. AFM results (Figure 1C) further illustrate that the topography of bio-imprinted POC-click membrane is comparable to the original tendon section template in submicron scale. These results reveal that the POC-click membrane can accurately duplicate the surface topography and physical dimension of natural tendon section at sub-micron levels. H&E staining images(Figure 2A) exhibit that MSCs cultured on POC-click membrane are elongated and aligned along the imprinted pattern on POC-click membrane. IHC staining (Figure 2B) illustrates that the tenogenic behavior of MSCs cultured on POC-click membrane is also comparable to the MSCs cultured on the natural bovine tendon section. Conclusion: Our bio-engineered POC-click membrane can faithfully replicate the TME of tendon and the MSCs can be then efficiently differentiated into tenocytes. We believe this polymer is promising to bridge the gap between our experiment model and clinical application.
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