Acid Phosphatase Activity In Osteoclasts Research Articles

Porous gelatin/tricalcium phosphate/genipin composites containing lumbrokinase for bone repair

Bone cell activities are very important in bone remodeling. This study investigates the effects of lumbrokinase on bone cell activities in cultures. Moreover, a biodegradable composite (GGT) containing genipin-crosslinked gelatin and β-tricalcium phosphate was prepared to carry lumbrokinase (GGTLK). Rat calvarial bone defects were filled with GGT and GGTLK composites. Bone healing was monitored in vivo by bioluminescence imaging and micro-CT. Lumbrokinase was found to have a dose-dependent effect on bone cell activities. Low concentrations (<1μg/ml) of lumbrokinase increased the viability, total alkaline phosphatase activity and mobility of osteoblasts, the number of total calcified nodules and the expression of osteopontin and osteocalcin; however, they considerably reduced the total tartrate-resistant acid phosphatase activity of osteoclasts. IVIS images revealed a stronger fluorescent signal in GGTLK-treated animals than in GGT-treated animals. Micro-CT analysis revealed that GGTLK induced more new bone formation than did GGT. These observations suggest that lumbrokinase released from GGTLK composite can enhance bone tissue regeneration.

Effect of water extract and distillate from the mixture of black goat meat and medicinal herb on osteoblast proliferation and osteoclast formation

Purpose: The effects of water extract and distillate from the mixture of black goat meat and medicinal herb on MG-63 osteoblast proliferation and mouse bone marrow derived osteoclast formation were investigated. Methods: Proximate composition, volatile basic nitrogen (VBN), mineral content, free amino acid composition and free fatty acid composition in black goat meat were determined. Water extract and distillate were prepared with three groups; goat meat only (BG-E, BGD), six herbs added group (BG-E6, BG-D6), and eight herbs added group (BG-E8, BG-D8). Osteoblast proliferation, mineralization and calcium uptake activity of MG-63 cells were measured and tartrate resistant acid phosphatase activity of osteoclasts was analyzed. Results: Black goat meat had remarkably low fat and high level of calcium. Glutamic acid was the most abundant amino acid. Herbs added extract groups (BG-E6 and BG-E8) showed increased MG-63 cell proliferation in a concentration dependent manner, while all the distillates did not show the effect. All extracts and distillates showed significantly increased osteoblast mineralization depending on the concentration. In particular, herb added extract, BG-E6, increased 170.3% of control and the distillate of BG-D and BG-D6 increased up to 168.5% and 159.8%, respectively. Calcium uptake activities of all water extracts showed remarkable increase of BG-E6 and BG-E8 up to 615.5% and 628.1% of control, respectively. Ditillates had no effect except BG-D6. All water extracts significantly reduced the activity of tartrate-resistant acid phosphatase (TRAP) in osteoclasts derived from mouse bone marrow. Conclusion: Combination of black goat meat and medicinal herb increased the MG-63 cell proliferation and effectively inhibited osteoclast differentiation in both water extracts and distillate of them, which implies that they could be used as potent functional food materials for bone health.

Earthworm (Pheretima aspergillum) extract stimulates osteoblast activity and inhibits osteoclast differentiation.

BackgroundThe potential benefits of earthworm (Pheretima aspergillum) for healing have received considerable attention recently. Osteoblast and osteoclast activities are very important in bone remodeling, which is crucial to repair bone injuries. This study investigated the effects of earthworm extract on bone cell activities.MethodsOsteoblast-like MG-63 cells and RAW 264.7 macrophage cells were used for identifying the cellular effects of different concentrations of earthworm extract on osteoblasts and osteoclasts, respectively. The optimal concentration of earthworm extract was determined by mitochondrial colorimetric assay, alkaline phosphatase activity, matrix calcium deposition, Western blotting and tartrate-resistant acid phosphatase activity.ResultsEarthworm extract had a dose-dependent effect on bone cell activities. The most effective concentration of earthworm extract was 3 mg/ml, significantly increasing osteoblast proliferation and differentiation, matrix calcium deposition and the expression levels of alkaline phosphatase, osteopontin and osteocalcin. Conversely, 3 mg/ml earthworm extract significantly reduced the tartrate-resistant acid phosphatase activity of osteoclasts without altering cell viability.ConclusionsEarthworm extract has beneficial effects on bone cell cultures, indicating that earthworm extract is a potential agent for use in bone regeneration.

Tetracycline administration normalizes the structure and acid phosphatase activity of osteoclasts in streptozotocin‐induced diabetic rats

Diabetes induces osteopenia, which is characterized by a deficiency of osteoid and decreased activity of osteoblasts. We recently found that tetracyclines prevent the loss of osteoid and bone matrix and the degeneration of osteoblasts in diabetic rats by a mechanism independent of their antimicrobial efficacy. However, bone remodeling requires the activity of osteoclasts as well as osteoblasts. To determine the in vivo effects of tetracycline on osteoclasts in long bones, either a tetracycline (minocycline, TC) or its chemically modified non-antibiotic analogue (CMT), 4-de-dimethylaminotetracycline, was administrated daily to streptozotocin-induced diabetic rats by oral intubation. After 21 days, the rats were perfusion-fixed with a mixture of formaldehyde and glutaraldehyde, and the humeri were dissected and processed for ultracytochemical demonstration of acid trimetaphosphatase (ACPase) activity. In untreated non-diabetic (control) rats, the osteoclasts at the zone of provisional ossification exhibited abundant mitochondria and cisterns of rough endoplasmic reticulum (RER) throughout the cytoplasm, prominent stacks of Golgi membranes, and lysosomes in the perinuclear cytoplasm, and numerous various pale vacuoles in the cytoplasmic area adjacent to well-developed ruffled border. Intense ACPase activity was observed in the Golgi saccules, lysosomes, pale vacuoles, and the extracellular canals of ruffled border. The reaction products were also noted along the resorbing bone surfaces associated with the osteoclast ruffled border. The osteoclasts in the untreated diabetic rats showed a cytoplasmic organization similar to that of the non-diabetic control rats, but showed little or no ruffled border which was replaced by a broad clear zone in some of these cells. However, most of the osteoclasts on bone matrix in the diabetics were devoid of both a ruffled border and a clear zone. ACPase activity was detected in the osteoclast cytoplasm of diabetic rat, as in the controls, but to a much lesser extent along the broad clear zone facing the resorbing bone surfaces. The osteoclasts in TC-treated diabetic rats possessed both a clear zone and a small ruffled border. However, in some cases, they lacked both structures reminiscent of the untreated diabetic cells. The osteoclasts of CMT-treated diabetic rats exhibited structural and enzymatic features essentially identical to those of the non-diabetic control rats. These results suggest that the diabetes-induced osteopenia results, at least in part, from degeneration of osteoclasts (as well as atrophic osteoblasts) and that tetracyclines may be effective in preventing these abnormalities by a mechanism not dependent on the drugs' antimicrobial properties.

Osteoarthrosis

The present study was undertaken to investigate the sites at which the 99mTc phosphorous compounds bind to skeletal tissue in general, and their localization at different stages of osteoarthritis in particular, in order thereby to arrive at a better morphological basis for interpreting the scintigrams. It was endeavoured also to relate the uptake of bone-seeking agents to abnormal changes in cartilage, synovium, and subchondral bone to obtain better insight into the pathogenesis of osteoarthritis. The bone remodelling activities in subchondral bone in osteoarthritic human femoral heads were elucidated by the alkaline phosphatase activity of osteoblasts and the acid phosphatase activity of osteoclasts. The enzyme activity was measured semiquantitatively by the initial time for the histochemical reaction. The distribution of the activity of the two enzymes in different areas proved parallel, and considerable variation in enzyme activity was seen between different areas within the same femoral head. Increased osteoarthritic cartilaginous changes were associated with increased subchondral enzyme activity, highest in denuded weightbearing areas and in the osteophytes and lowest in non-weightbearing subchondral bone and centrally in the femoral head. Studies on different histochemical staining of glycosaminoglycans in the matrix of human osteoarthritic cartilage and of normal cartilage revealed a heterogeneous distribution of the different glycosaminoglycans through the cartilage. Except for superficial loss of glycosaminoglycans, no difference was found in the distribution of keratan sulphate between osteoarthritic cartilage and control cartilage. In osteoarthritis, however, a relative increase in stainability for chondroitin sulphate was found in the territorial area, especially around the cell clusters, and only chondroitin sulphate was present in the cartilage of osteophytes. These findings were interpreted as an increased GAG metabolism, its mode of production being like that of very young cartilage. In the experimental rabbit model used in studying the uptake of bone-seeking agents this GAG regeneration was able to refill demasked collagen network with glycosaminoglycans in certain areas of the joint. The height of the depleted superficial area was estimated on patellar cartilage stained for sulphated GAG with toluidine blue-0 at pH 3, visually and by optical densitometry using the wavelength corresponding to the gamma-band of toluidine blue. The time relation of the surface depletion was elucidated. A marked depletion of GAG, seen one week postoperatively, reached a maximum at 4 weeks.(ABSTRACT TRUNCATED AT 400 WORDS)

A quantitative cytochemical assay for acid phosphatase activity in osteoclasts in the foetal rat calvaria

A quantitative cytochemical assay for acid phosphatase activity in osteoclasts in the foetal rat calvaria

Alkaline and Acid Phosphatase Activity in Osteoclasts, Osteoblasts and Giant Cell Tumors of Bone An Ultrastructural, Cytochemical Study

Alkaline and Acid Phosphatase Activity in Osteoclasts, Osteoblasts and Giant Cell Tumors of Bone An Ultrastructural, Cytochemical Study