Measurement of Total Alkaline Phosphatase Activity in Human Serum

Bone engineering opens the possibility to grow large amounts of tissue products by combining patient-specific cells with compliant biomaterials. Decellularized tissue matrices represent suitable biomaterials, but availability, long processing time, excessive cost, and concerns on pathogen transmission have led to the development of biomimetic synthetic alternatives. We recently fabricated calcium phosphate cement (CPC) scaffolds with variable macroporosity using a facile synthesis method with minimal manufacturing steps and demonstrated long-term biocompatibility in vitro. However, there is no knowledge on the potential use of these scaffolds for bone engineering and whether the porosity of the scaffolds affects osteogenic differentiation and tissue formation in vitro. In this study, we explored the bone engineering potential of CPC scaffolds with two different macroporosities using human mesenchymal progenitors derived from induced pluripotent stem cells (iPSC-MP) or isolated from bone marrow (BMSC). Biomimetic decellularized bone scaffolds were used as reference material in all experiments. The results demonstrate that, irrespective of their macroporosity, the CPC scaffolds tested in this study support attachment, viability, and growth of iPSC-MP and BMSC cells similarly to decellularized bone. Importantly, the tested materials sustained differentiation of the cells as evidenced by increased expression of osteogenic markers and formation of a mineralized tissue. In conclusion, the results of this study suggest that the CPC scaffolds fabricated using our method are suitable to engineer bone grafts from different cell sources and could lead to the development of safe and more affordable tissue grafts for reconstructive dentistry and orthopaedics and in vitro models for basic and applied research.

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“…Alkaline phosphatase (ALP) activity was determined using a modification of the method described in the literature (Bowers & McComb, 1975…”

Section: Alkaline Phosphatase Activitymentioning

confidence: 99%

Engineering human bone grafts with new macroporous calcium phosphate cement scaffolds

Sladkova

Palmer

Öhman

et al. 2017

J Tissue Eng Regen Med

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“…An aliquot of blood serum previously collected from each family member was assessed for phosphorous and calcium by the clinical laboratories of the University of Washington Hospital using standard techniques, and the level of alkaline phosphatase activity was measured by the method of Bowers & McComb (1975). Urinary phosphoethanolamine (PEA) levels were measured on a Durham amino acid analyzer using standard methods for analysis of physiologic fluids, and the content calculated on the basis of previously tested standards.…”

Section: Blood and Urine Analysismentioning

confidence: 99%

Laboratory studies of a family manifesting premature exfoliation of deciduous teeth

Baab

Page

Ebersole

et al. 1986

J Clinic Periodontology

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In a previous publication, we described the clinical and radiographic findings of a family in which the children manifested premature exfoliation of the deciduous teeth. We now report for the same family the results of extensive laboratory studies performed on blood and urine, analysis of periodontal microflora, and a family pedigree. We demonstrated the presence of putative periodontal pathogens in the subgingival microflora, elevated levels of serum antibodies reacting to Bacteroides gingivalis, Capnocytophaga gingivalis, and C. sputigena in 2 of the children, and significantly suppressed monocyte chemotaxis in all 3 children. Phosphoethanolamine was found in the urine of the father and all 3 children, but not in the mother. Likewise, serum alkaline phosphatase was abnormally low for all 3 children, and was at the extreme low end of normal range for the father, but was normal for the mother. On the basis of the alkaline phosphatase and phosphoethanolamine measurements, we assigned a diagnosis of hypophosphatasia to the 3 children. Phosphoethanolamine and alkaline phosphatase were also abnormal in the paternal grandmother and her brother. The son of this brother who was deceased had a daughter manifesting premature loss of the primary teeth. The data are consistent with an autosomal dominant mode of transmission. In the light of our findings, hypoplastic cementum must be considered in the etiology of some forms of early-onset periodontitis.

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“…The analysis of AcP and PAP were performed as previously described method of Bowers and McComb [18]. The final concentration of the reactants were diethanolamine, 900 mM; MgC12, 0.1 mM; p-nitrophenyl phosphate, 10 mM; and a serum dilution of 1: 126.…”

Section: Experimental Methodsmentioning

confidence: 99%

The use of serum isoenzymes of alkaline and acid phosphatase as possible quantitative markers of tumor load in prostate cancer

et al. 1981

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The tumor burden of 98 patients with metastatic prostatic cancer was compared longitudinally with the activities of bone (BAP) and liver isoenzymes (LAP) of alkaline phosphatase, total acid phosphatase (AcP), and prostate-specific acid phosphatase (PAP). A quantitative association between these enzyme markers and the tumor mass was suggested by comparing the enzymes with 1) both the treatment response and the estimation of metastasis by radionuclide bone scanning; 2) metastasis based upon radiographic evidence. In addition, an apparent extensive pretreatment bone tumor load was predictive for an elevated BAP activity, which was also a suggestive poor prognosis as previously reported. An elevation of PAP, in contrast to AcP, may precede the clinical disease progression in some patients. Data presented in this report have indicated that the levels of these enzymes compared well with the extent of tumor involvement and therefore may be considered suitable as adjuvant and even quantitative biochemical markers of bone and liver metastasis.

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Measurement of Total Alkaline Phosphatase Activity in Human Serum

Cited by 213 publications

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Engineering human bone grafts with new macroporous calcium phosphate cement scaffolds

Engineering human bone grafts with new macroporous calcium phosphate cement scaffolds

Laboratory studies of a family manifesting premature exfoliation of deciduous teeth

The use of serum isoenzymes of alkaline and acid phosphatase as possible quantitative markers of tumor load in prostate cancer

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