{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T22:18:12Z","timestamp":1788301092808,"version":"build-2803163510"},"reference-count":57,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2018,5,28]],"date-time":"2018-05-28T00:00:00Z","timestamp":1527465600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Gelatins from the skin of bovine, porcine, and tilapia were hydrolyzed to three degrees of hydrolysis (DH) by alcalase, neutrase, and papain, respectively. These hydrolysates at 0.02\u20130.1 g\/L promoted the growth of human fetal osteoblasts by 101.4\u2013135.7%, while higher DH or using papain and tilapia gelatins resulted in higher proliferation. The hydrolysates from porcine and tilapia gelatins at 0.05 g\/L prevented induced apoptosis (decreasing total apoptotic proportions from 28.4% or 35.2% to 10.3\u201317.5% or 16.0\u201323.6%), and had differentiation induction (increasing alkaline phosphatase activity by 126.9\u2013246.7% in early differentiation stage, or enhancing osteocalcin production by 4.1\u201322.5% in later differentiation stage). These hydrolysates had a similar amino acid profile; however, tilapia gelatin hydrolysates by papain with DH 15.4% mostly displayed higher activity than others. Tilapia gelatin hydrolysate could up-regulate \u03b2-catenin, Wnt 3a, Wnt 10b, cyclin D1, and c-Myc expression at mRNA levels by 1.11\u20133.60 folds, but down-regulate GSK 3\u03b2 expression by 0.98 fold. Of note, \u03b2-catenin in total cellular and nuclear protein was up-regulated by 1.14\u20131.16 folds but unchanged in cytoplasmic protein, Wnt 10b, cyclin D1, and c-Myc expression were up-regulated by 1.27\u20131.95 folds, whilst GSK 3\u03b2 expression was down-regulated by 0.87 fold. Activation of Wnt\/\u03b2-catenin pathway is suggested to mediate cell proliferation and differentiation.<\/jats:p>","DOI":"10.3390\/molecules23061287","type":"journal-article","created":{"date-parts":[[2018,5,29]],"date-time":"2018-05-29T02:58:18Z","timestamp":1527562698000},"page":"1287","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["The Growth Proliferation, Apoptotic Prevention, and Differentiation Induction of the Gelatin Hydrolysates from Three Sources to Human Fetal Osteoblasts (hFOB 1.19 Cells)"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5069-6665","authenticated-orcid":false,"given":"Ming","family":"Lu","sequence":"first","affiliation":[{"name":"Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9682-5426","authenticated-orcid":false,"given":"Xin-Huai","family":"Zhao","sequence":"additional","affiliation":[{"name":"Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China"},{"name":"Department of Food Science, Northeast Agricultural University, Harbin 150030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,5,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"25103","DOI":"10.1074\/jbc.R109.041087","article-title":"Cellular and molecular mechanisms of bone remodeling","volume":"285","author":"Raggatt","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ejphar.2014.01.013","article-title":"Chlormadinone acetate promotes osteoblast differentiation of human mesenchymal stem cells through the ERK signaling pathway","volume":"726","author":"Kim","year":"2014","journal-title":"Eur. J. Pharmacol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1508","DOI":"10.1126\/science.289.5484.1508","article-title":"Therapeutic approaches to bone diseases","volume":"289","author":"Rodan","year":"2000","journal-title":"Science"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1575","DOI":"10.1556\/OH.2014.30015","article-title":"Bone tissue: Rebuilding and inflammation","volume":"155","author":"Jakab","year":"2014","journal-title":"Orvosi Hetil."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.phrs.2015.04.004","article-title":"Ginkgo biloba extract promotes osteogenic differentiation of human bone marrow mesenchymal stem cells in a pathway involving Wnt\/\u03b2-catenin signaling","volume":"97","author":"Gu","year":"2015","journal-title":"Pharmacol. Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1016\/j.rdc.2006.08.003","article-title":"Epidemiology of osteoporosis","volume":"32","author":"Dennison","year":"2006","journal-title":"Rheum. Dis. Clin. N. Am."},{"key":"ref_7","first-page":"507","article-title":"Postmenopausal osteoporosis and hormone replacement therapy","volume":"95","author":"Gambacciani","year":"2004","journal-title":"Minerva Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"869","DOI":"10.1016\/j.beem.2008.06.003","article-title":"Bone-forming agents in the management of osteoporosis","volume":"22","author":"Neuprez","year":"2008","journal-title":"Best Pract. Res. Clin. Endocrinol. Metab."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1007\/s00774-013-0424-2","article-title":"Non-adherence to anti-osteoporotic medications in Taiwan: Physician specialty makes a difference","volume":"31","author":"Yu","year":"2013","journal-title":"J. Bone Miner. Metab."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2028","DOI":"10.1056\/NEJMoa071408","article-title":"Teriparatide or alendronate in glucocorticoid-induced osteoporosis","volume":"357","author":"Saag","year":"2007","journal-title":"N. Engl. J. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"909","DOI":"10.3945\/ajcn.117.153353","article-title":"Combined bioavailable isoflavones and probiotics improve bone status and estrogen metabolism in postmenopausal osteopenic women: A randomized controlled trial","volume":"106","author":"Lambert","year":"2017","journal-title":"Am. J. Clin. Nutr."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"199","DOI":"10.3177\/jnsv.43.199","article-title":"Effects of whey protein on calcium and bone metabolism in ovariectomized rats","volume":"43","author":"Takada","year":"1997","journal-title":"J. Nutr. Sci. Vitaminol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1053\/sarh.2000.9622","article-title":"Role of collagen hydrolysate in bone and joint disease","volume":"30","author":"Moskowitz","year":"2000","journal-title":"Semin. Arthritis Rheum."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1093\/jn\/131.3.723","article-title":"Soybean isoflavones dose-dependently reduce bone turnover but do not reverse established osteopenia in adult ovariectomized rats","volume":"131","author":"Picherit","year":"2001","journal-title":"J. Nutr."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Schadow, S., Siebert, H.C., Lochnit, G., Kordelle, J., Rickert, M., and Steinmeyer, J. (2013). Collagen metabolism of human osteoarthritic articular cartilage as modulated by bovine collagen hydrolysates. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0053955"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1007\/s00774-004-0522-2","article-title":"Assessment of effectiveness of oral administration of collagen peptide on bone metabolism in growing and mature rats","volume":"22","author":"Wu","year":"2004","journal-title":"J. Bone Miner. Metab."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e35","DOI":"10.1186\/1550-2783-10-35","article-title":"Hydrolyzed collagen intake increases bone mass of growing rats trained with running exercise","volume":"10","author":"Takeda","year":"2013","journal-title":"J. Int. Soc. Sports Nutr."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1120","DOI":"10.1016\/j.nut.2005.03.007","article-title":"Increase in bone mineral density through oral administration of shark gelatin to ovariectomized rats","volume":"21","author":"Nomura","year":"2005","journal-title":"Nutrition"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2680","DOI":"10.1271\/bbb.70287","article-title":"Promotion by collagen tripeptide of type I collagen gene expression in human osteoblastic cells and fracture healing of rat femur","volume":"71","author":"Tsuruoka","year":"2007","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/j.jff.2012.10.017","article-title":"In vitro responses of hFOB 1.19 cells towards chum salmon (Oncorhynchus keta) skin gelatin hydrolysates in cell proliferation, cycle progression and apoptosis","volume":"5","author":"Fu","year":"2013","journal-title":"J. Funct. Foods"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1016\/j.foodchem.2017.09.043","article-title":"Blue mussel (Mytilus edulis) protein hydrolysate promotes mouse mesenchymal stem cell differentiation into osteoblasts through up-regulation of bone morphogenetic protein","volume":"242","author":"Hyung","year":"2018","journal-title":"Food Chem."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Liu, J.L., Zhang, B., Song, S.J., Ma, M., Si, S.Y., Wang, Y.H., Xu, B.X., Feng, K., Wu, J.G., and Guo, Y.C. (2014). Bovine collagen peptides compounds promote the proliferation and differentiation of MC3T3-E1 pre-osteoblasts. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0099920"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"84","DOI":"10.3177\/jnsv.47.84","article-title":"Ingestion of gelatin has differential effect on bone mineral density and body weight in protein undernutrition","volume":"47","author":"Koyama","year":"2001","journal-title":"J. Nutr. Sci. Vitaminol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1016\/j.bone.2009.10.035","article-title":"Hydrolyzed collagen improves bone metabolism and biomechanical parameters in ovariectomized mice: An in vitro and in vivo study","volume":"46","author":"Guillerminet","year":"2010","journal-title":"Bone"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"15474","DOI":"10.3390\/molecules181215474","article-title":"Osteogenic activity of collagen peptide via ERK\/MAPK pathway mediated boosting of collagen synthesis and its therapeutic efficacy in osteoporotic bone by back-scattered electron imaging and microarchitecture analysis","volume":"18","author":"Kim","year":"2013","journal-title":"Molecules"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1532","DOI":"10.1021\/jf062834s","article-title":"Comparison of quantity and structures of hydroxyproline-containing peptides in human blood after oral ingestion of gelatin hydrolysates from different sources","volume":"55","author":"Ohara","year":"2007","journal-title":"J. Agric. Food Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1016\/j.bbrc.2004.11.072","article-title":"Regulation of apoptosis in osteoclasts and osteoblastic cells","volume":"328","author":"Xing","year":"2005","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"793","DOI":"10.1359\/jbmr.1998.13.5.793","article-title":"Osteoblast programmed cell death (apoptosis): Modulation by growth factors and cytokines","volume":"13","author":"Jilka","year":"1998","journal-title":"J. Bone Miner. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1132","DOI":"10.1038\/nm1096-1132","article-title":"Estrogen promotes apoptosis of murine osteoclasts mediated by TGF-\u03b2","volume":"2","author":"Hughes","year":"1996","journal-title":"Nat. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4632","DOI":"10.1074\/jbc.M411530200","article-title":"Transient versus sustained phosphorylation and nuclear accumulation of ERKs underlie anti-versus pro-apoptotic effects of estrogens","volume":"280","author":"Chen","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_31","first-page":"719","article-title":"Nongenotropic, sex-nonspecific signaling through the estrogen or androgen receptors: Dissociation from transcriptional activity","volume":"104","author":"Kousteni","year":"2001","journal-title":"Cell"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Wang, S., Fu, Y., and Zhao, X.H. (2016). The cooperative effect of genistein and protein hydrolysates on the proliferation and survival of osteoblastic cell (hFOB 1.19). Molecules, 21.","DOI":"10.3390\/molecules21111489"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.yexcr.2016.11.013","article-title":"Constitutive \u03b2-catenin activation in osteoblasts impairs terminal osteoblast differentiation and bone quality","volume":"350","author":"Bao","year":"2017","journal-title":"Exp. Cell Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.jnutbio.2016.08.003","article-title":"Akt drives buffalo casein-derived novel peptide-mediated osteoblast differentiation","volume":"38","author":"Reddi","year":"2016","journal-title":"J. Nutr. Biochem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1272\/jnms.77.4","article-title":"The mechanism of mineralization and the role of alkaline phosphatase in health and disease","volume":"77","author":"Orimo","year":"2010","journal-title":"J. Nippon Med. Sch."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e846480","DOI":"10.1155\/2013\/846480","article-title":"Endocrine actions of osteocalcin","volume":"2013","author":"Patti","year":"2013","journal-title":"Int. J. Endocrinol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"422","DOI":"10.1248\/bpb.35.422","article-title":"Distribution of prolylhydroxyproline and its metabolites after oral administration in rats","volume":"35","author":"Kawaguchi","year":"2012","journal-title":"Biol. Pharm. Bull."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1620","DOI":"10.1016\/j.joca.2009.07.001","article-title":"Chondroprotective effect of the bioactive peptide prolyl-hydroxyproline in mouse articular cartilage in vitro and in vivo","volume":"17","author":"Nakatani","year":"2009","journal-title":"Osteoarthr. Cartil."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"13908","DOI":"10.3390\/ijms160613908","article-title":"In vitro proliferation and anti-apoptosis of the papain-generated casein and soy protein hydrolysates towards osteoblastic cells (hFOB1.19)","volume":"16","author":"Pan","year":"2015","journal-title":"Int. J. Mol. Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1101\/gad.1394306","article-title":"CKI, there\u2019s more than one: Casein kinase I family members in Wnt and Hedgehog signaling","volume":"20","author":"Price","year":"2006","journal-title":"Genes Dev."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"751","DOI":"10.1016\/j.devcel.2005.02.017","article-title":"Canonical Wnt signaling in differentiated osteoblasts controls osteoclast differentiation","volume":"8","author":"Glass","year":"2005","journal-title":"Dev. Cell"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"21162","DOI":"10.1074\/jbc.M501900200","article-title":"Essential role of beta-catenin in postnatal bone acquisition","volume":"280","author":"Holmen","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1016\/S1359-6101(00)00011-3","article-title":"Wnt\/\u03b2-catenin signaling","volume":"11","author":"Akiyama","year":"2000","journal-title":"Cytokine Growth Factor Rev."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1080\/08977190412331279890","article-title":"Bone morphogenetic proteins","volume":"22","author":"Chen","year":"2004","journal-title":"Growth Factors"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1128\/MCB.19.1.1","article-title":"c-Myc target genes involved in cell growth, apoptosis, and metabolism","volume":"19","author":"Dang","year":"1999","journal-title":"Mol. Cell. Biol."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Mirando, A.J., Maruyama, T., Fu, J., Yu, H.M.I., and Hsu, W. (2010). Beta-catenin\/cyclin D1 mediated development of suture mesenchyme in calvarial morphogenesis. BMC Dev. Biol., 10.","DOI":"10.1186\/1471-213X-10-116"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1016\/j.bone.2009.10.024","article-title":"Osteo-transcriptomics of human mesenchymal stem cells: Accelerated gene expression and osteoblast differentiation induced by vitamin D reveals c-MYC as an enhancer of BMP2-induced osteogenesis","volume":"46","author":"Piek","year":"2010","journal-title":"Bone"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1359\/jbmr.081235","article-title":"Where Wnts went: The exploding field of Lrp 5 and Lrp 6 signaling in bone","volume":"24","author":"Williams","year":"2009","journal-title":"J. Bone Miner. Res."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1016\/j.semarthrit.2011.01.006","article-title":"The emerging role of Dickkopf-1 in bone biology: Is it the main switch controlling bone and joint remodeling?","volume":"41","author":"Daoussis","year":"2011","journal-title":"Semin. Arthritis Rheum."},{"key":"ref_50","first-page":"139","article-title":"Effect of estrogens and antiestrogen receptor dynamics and the induction of progesterone receptor in MCF-7 human breast cancer cells","volume":"42","author":"Eckert","year":"1982","journal-title":"Cancer Res."},{"key":"ref_51","unstructured":"Association of Official Agricultural Chemists (AOAC) (2005). Official Methods of Analysis of Association of Official Analytical Chemists International, AOAC International. [18th ed.]."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1961","DOI":"10.1021\/ac60205a053","article-title":"Two improved and simplified methods for the spectrophotometric determination of hydroxyproline","volume":"35","author":"Bergman","year":"1963","journal-title":"Anal. Chem."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"178","DOI":"10.1002\/jbmr.5650100203","article-title":"Development and characterization of a conditionally immoralized human fetal osteoblastic cell line","volume":"10","author":"Harris","year":"1995","journal-title":"J. Bone Miner. Res."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/j.jconrel.2014.06.029","article-title":"Osteogenic differentiation of human mesenchymal stem cells through alginate-graft-poly (ethylene glycol) microsphere-mediated inracellular growth factor delivery","volume":"192","author":"Miao","year":"2014","journal-title":"J. Control. Release"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/0022-1759(95)00072-I","article-title":"A novel assay for apoptosis Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V","volume":"184","author":"Vermes","year":"1995","journal-title":"J. Immunol. Methods"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2212\u2206\u2206Ct method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1016\/j.yexcr.2017.12.006","article-title":"K-ras mutation promotes ionizing radiation-induced invasion and migration of lung cancer in part via the Cathepsin L\/CUX1 pathway","volume":"362","author":"Wang","year":"2018","journal-title":"Exp. Cell Res."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/23\/6\/1287\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:06:09Z","timestamp":1760195169000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/23\/6\/1287"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,5,28]]},"references-count":57,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2018,6]]}},"alternative-id":["molecules23061287"],"URL":"https:\/\/doi.org\/10.3390\/molecules23061287","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,5,28]]}}}