Cartilage tissue engineering in the dynamic culture under hypoxic conditions

Background:

The culture of human chondrocytes (hPCh) on decellularized porcine cartilage scaffolds (DECM) in a bioreactor vessel (BR) showed a very good migration of the hPCh into the DECM under normoxic conditions but demonstrated a low potential of re-differentiation. As cartilage has no direct blood supply chondrocytes are poorly supplied with oxygen. We therefore hypothesized that based on the physical conditions the cell differentiation under hypoxic conditions would probably be superior to that under normoxic conditions.

Material and methods:

DECM seeded with hPCh are cultured in a specially developed BR under hypoxic conditions and removed after three defined time periods. The static culture serves as control whereby the DECM are cultured horizontally and vertically. The cartilage constructs are histologically and immunohistochemically analyzed and the matrix production is examined by PCR and DMMB-Assay.

Results:

The BR also supports the cell migration under hypoxic conditions. The production of cartilage specific proteins and glycosaminoglycans can be proven by PCR and DMMB-assay in both dynamic and static cultivation whereby the re-differentiation proceeds much slower in the BR. In comparison to the cultivation under normoxia all of the expression levels are lower. The vertical static cultivation is superior to the horizontal in all examined parameters.

Conclusion:

Even in the culture under hypoxic conditions we find a good cell migration and matrix synthesis. However, the differentiation of the hPCh is not speeded up under hypoxia. The influence of other factors on the differentiation behavior of the hPCh must be examined in further studies.

Publication History

Publication Date:
18 April 2018 (online)