Differentiation and Anti-inflammatory Potentials of Eucomis autumnalis and Pterocarpus angolensis Extracts Scaffolds in Porcine Adipose–Derived Mesenchymal Stem Cells

Abstract

The key ingredients for tissue engineering and regenerative medicine are type of cells, growth factors, and biological engineering materials/scaffolds. Primary porcine adipose–derived mesenchymal stem cells (pADMSCs) were characterized using fluorescence-activated cell sorting analysis for mesenchymal surface markers of CD44, CD90, and CD105 (≥\u200985%). Cell attachment was evaluated using scanning electron microscope (SEM) and immunofluorescence. MTT and histology assays were employed to evaluate pADMSCs viability and multi-potency differentiation respectively on Eucomis autumnalis and Pterocarpus angolensis crude extracts scaffolds. IL-6 ELISA and RT-PCR were used to access anti-inflammatory and bone/cartilage marker expressions respectively. Results showed that multi-potency differentiation of pADMSCs was high and dose dependent on groups treated with E. autumnalis and P. angolensis. SEM and immunofluorescence results revealed more cells attached after 7, 14, and 21 days on treated groups than untreated groups. Cell viability was high with significant difference of P <\u20090.01 in plant extract groups after 7, 14, and 21 days. The E. autumnalis crude extract significantly (P <\u20090.01) suppressed the protein expression of IL-6 in comparison to extracts prepared from P. angolensis and untreated groups. Collagen type II and osteopontin were significantly expressed in P. angolensis group compared to E. autumnalis and other groups. Alkaline phosphatase and osteocalcin genes were significantly expressed on days 14 and 21 in E. autumnalis compared to other groups. This study provides some evidence to support the anti-inflammatory nature of E. autumnalis and P. angolensis extract as well as proof that these plants are able to induce multi-potent differentiation of ADMSCs. Eucomis autumnalis and Pterocarpus angolensis are among the plants used by numerous traditional healers to reduce osteoarthritis symptoms and improve bone fracture healings. This work explored the potentials of plant extracts incorporated scaffolds to induce adipogenic, osteogenic, chondrogenic differentiation, and anti-inflammatory effects in porcine adipose-derived mesenchymal stem cells (pADMSCs). Our data showed that plant extract scaffolds suppressed the expression of interlukin-6 especially in E. autumnalis in our ELISA assay analysis which confirmed the anti-inflammatory nature of the plant extract scaffolds. The plant extract scaffolds also depicted osteogenic and chondrogenic support in the multi-potency differentiation of the pADMSCs. Pterocarpus angolensis extract scaffold showed high upregulated expression of collagen type II on day 21 at the highest dose of 5 mg/ml. In conclusion, plant extracts in scaffolds may provide safe, cost-effective alternative treatment for bone healing and cartilage regeneration and stimulation of bone formation by virtue of their anti-inflammatory properties. Our research work is relevant to communities that rely solely on traditional medicine for their well-being, orthopedics, pharmaceutical companies, regenerative, and translational medicine fields.