Enzymatic Cell Isolation and Explant Cultures of Rat Calvarial Osteoblast Cells

Osteoblast cells were isolated from the calvarial bones of newborn Wistar rats and cultured in vitro via both collagenase digestion method and explant technique, and a comparative study was carried out on the two culture methods. The biologic charwteristics of the osteoblast cells were studied via cell number counting, morphology observation, alkaline phosphatase staining of the cells and alizarine- red staining of the calcified nodules. The results show that osteoblast cells can be cultured in vitro via collagenase digestion method and explant technique, and the obtained cells ure of good biologic characteristics. In comparison with the explant technique, the operative procedure of the enzymatic digestion method is more complicated. The digestion time must be carefully controlled. However, with this method, one can obtain a lager number of cells in a short time. The operative procedure of the explant technique is simpler, but it usually takes longer time to obtain cells of desirable number.

Activity of the Mechanically Isolated Hybrid Stromal Vascular Fraction on Osteoarthritis

Background: Regenerative medicine holds promise for treating degenerative and inflammatory conditions like osteoarthritis (OA). However, the complex molecular mechanisms of OA and the limitations of current therapies remain challenges. Adipose-derived stem cells (ADSC) and stromal vascular fraction (SVF) are gaining attention for OA treatment due to their abundance in adipose tissue. The literature delineates two primary techniques for SVF extraction from adipose tissue: enzymatic digestion and mechanical methods. The Lipocube Hybrid SVF presents a straightforward and secure mechanical isolation method for SVF, enhancing its regenerative potential for various applications. Purpose: This study aims to provide valuable insights into the potential of Lipocube Hybrid SVF as a regenerative therapy for OA, contributing to the broader understanding of its applicability in addressing this debilitating condition. Method: To assess the effectiveness and safety of the Lipocube Hybrid SVF, we have designed a comparative study that evaluates cellular activity and viability, phenotypic characterization, and differentiation potential. The in vitro activity of mechanically isolated SVF is compared to the established gold standard enzymatic digestion method. After in vitro studies, Lipocube Hybrid mechanical isolation method was used to isolate SVF and applied in 42 knee and 7 hip joints of 28 patients with Grade II, Grade III, and Grade IV OA. Results: The Lipocube Hybrid group had slightly lower viable cell numbers but higher cell viability. Flow cytometry analysis showed the Lipocube Hybrid group exhibited more favorable markers for regenerative potential and reduced inflammatory response. Additionally, both groups demonstrated successful osteogenic differentiation, with the Lipocube Hybrid group excelling in chondrogenic and adipogenic differentiation. The clinical application of the Lipocube Hybrid SVF in OA patients resulted in significant improvements in WOMAC and VAS scores across different OA grades. Conclusions: This comparative study was conducted to evaluate the effectiveness and safety of the Lipocube Hybrid SVF, which has shown promise in laboratory settings, for different stages of osteoarthritis. The study findings provide valuable insights into the potential of Lipocube Hybrid SVF as a regenerative therapy for OA, highlighting its suitability for addressing this debilitating condition.

Evidence of Casparian Strip in the Foliar Endodermis of Pinus bungeana

利用荧光显微镜对白皮松 (PinusbungeanaZucc.)针叶内皮层结构进行深入观察 ,确认内皮层径向壁和横向壁上具有能够激发荧光的带状结构 ;用纤维素酶和果胶酶对内皮层充分酶解分离后 ,首次从叶中得到类似于根部内皮层特有的网状结构 ;应用傅立叶变换红外光谱分光术 (FTIR)对网状结构进行化学成分鉴定 ,其吸收光谱的分析结果表明 ,针叶内皮层细胞壁带状结构部分含有木质素、木栓质、纤维素和细胞壁蛋白等 ,而这些成分与针叶维管组织的成分不尽相同。上述证据表明 ,白皮松针叶内皮层细胞壁具有与根部内皮层相似的凯氏带结构。