PLATELET-RICH PLASMA ENRICHED CARDIAC EXTRACELLULAR MATRIX HYDROGEL INDUCES H9c2 CARDIOMYOBLAST PROLIFERATION AND DIFFERENTIATION
Name: EZIO HENRIQUE DA SILVA GOMES
Publication date: 16/02/2024
Examining board:
Name |
Role |
|---|---|
| BRENO VALENTIM NOGUEIRA | Advisor |
Summary: Injectable extracellular matrix (ECM)-based hydrogels are a prospective tool to be explored as a minimally invasive procedure and for cardiac tissue repair. The cardiac extracellular matrix (cECM) hydrogel is one the most promising injectable hydrogels. The use of hydrogels becomes more advantageous as it allows the incorporation of different substances and molecules of interest. Our group has used platelet-rich plasma, due to its easily accessible source of growth factors. Considering the aforementioned, we aimed to develop a Platelet Rich Plasma Cardiac Extracellular Matrix Hydrogel. To achieve this objective, myocardium from porcine ventricles was minced and decellularized in a 1% wt/vol sodium dodecyl sulfate (SDS) in 1× phosphate buffered saline (PBS) for 5-6 days, extensively washed with distilled water, frozen in an ultrafreezer at -80°C and lyophilized. Following, the material was enzymatically digested with pepsin and its pH adjusted to 7.4. Then the PRP was activated, using 330µL calcium chloride (100mg/mL) in 10mL of PRP. Ultimately, the activated PRP was incorporated into the gel in two concentrations: 10% and 20% (v/v). The materials were analyzed to assess residual DNA, histological analysis, SEM, FTIR, collagen content and biocompatibility in H9c2 cardiomyoblasts. After the process of decellularization the remaining DNA decreased from 758 (native) to 33 ng/mg dry tissue and the histological analysis showed absence of nuclei in the tissue, demonstrating the success of this process and reducing the chances of rejection reactions. The histochemistry and collagen content showed preservation of the collagen in the scaffold after the decellularization process. This preservation of collagen was critical for gel formation, allowing it to form a nanofibrous scaffold, seen by scanning electron microscopy. The encapsulation of H9c2 cardiomyoblast in hydrogel induced differentiation and the supplementation with PRP induce H9c2 cardiomyoblast proliferation. The results obtained in this work indicate that the injection of platelet-rich plasma enriched cardiac extracellular matrix hydrogel would help cardiac tissue repair after MI, once myoblasts would induce higher proliferationand differentiation.
