A Macroporous Bioreactor Super Activated by the Recombinant Human Transforming Growth Factor-β3

Abstract

Macroporous single phase hydroxyapatite (HA) and biphasic HA/β-tricalcium phosphate with 33% post-sinter hydroxyapatite (HA/β-TCP) were combined with 25 or 125 μg recombinant human transforming growth factor-β3 (hTGF-β3) to engineer a super activated bioreactor implanted in orthotopic calvarial and heterotopic rectus abdominis muscle sites and harvested on day 30 and 90. Coral-derived calcium carbonate fully converted (100%) and partially converted to 5 and 13% hydroxyapatite/calcium carbonate (5 and 13% HA/CC) pre-loaded with 125 and 250 μg hTGF-β3, and 1:5 and 5:1 binary applications of hTGF-β3: hOP-1 by weight, were implanted in the rectus abdominis and harvested on day 20 and 30, respectively, to monitor spatial/temporal morphogenesis by high doses of hTGF-β3. Bone formation was assessed on decalcified paraffin-embedded sections by measuring the fractional volume of newly formed bone. On day 30 and 90, single phase HA implants showed greater amounts of bone when compared to biphasic specimens; 5 and 13% HA/CC pre-loaded with 125 and 250 μg hTGF-β3 showed substantial induction of bone formation; 250 μg hTGF-β3 induced as yet unreported massive induction of bone formation as early as 20 days prominently outside the profile of the macroporous constructs. The induction of bone formation is controlled by the implanted ratio of the recombinant morphogens, i.e., the 1:5 hTGF-β3:hOP-1 ratio by weight was greater than the inverse ratio. The unprecedented tissue induction by single doses of 250 μg hTGF-β3 resulting in rapid bone morphogenesis of vast mineralized ossicles with multiple trabeculations surfaced by contiguous secreting osteoblasts is the novel molecular and morphological frontier for the induction of bone formation in clinical contexts.

Profiling bone morphogenetic proteins and transforming growth factor-βs by hTGF-β3 pre-treated coral-derived macroporous bioreactors: The power of one

“…Our previous studies showed the inhibitory effect of 125 mg hNoggin with or without 125 mg hTGF-b 3 on day 60 [2], so the present experiment focused on the morphological and now molecular analyses of the substantial induction of bone formation by the 250 mg doses of hTGF-b 3 , which were previously reported though without qRT-PCR analyses of the harvested specimens [15].…”

Cementogenesis and osteogenesis in periodontal tissue regeneration by recombinant human transforming growth factor‐β3: a pilot study in Papio ursinus

“…In each animal, two coral‐derived macroporous bioreactors super activated by 250 μ g hTGF‐ β 3 (Ripamonti et al. ,b) were implanted in the r. abdominis muscle as positive controls to correlate the induction of bone formation in the treated periodontal sites with de novo bone induction in heterotopic sites. Coral‐derived bioreactors solo without hTGF‐ β 3 were implanted as controls.…”

Redefining the induction of periodontal tissue regeneration in primates by the osteogenic proteins of the transforming growth factor‐β supergene family

“…Since the nineties, we have highlighted important challenges for the future of periodontal tissue regeneration ; the first challenge is still unresolved, i.e., “Which are the cell populations originating from both the periodontal ligament space and the residual bony housings that respond to the morphogenetic message of the soluble molecular signals of the TGF‐β supergene family?” . The second challenge, which we then thought to be the greatest challenge of all , has now re‐settled only to developmental events occurring in the normal course of embryonic development, as both marketed hOP‐1 and hBMP‐2 have been discontinued for lack of safety, limited effectiveness in clinical contexts, serious collateral effects and regulatory issues with the FDA .…”

“…(7,9). The second challenge, which we then thought to be the greatest challenge of all (7), has now re-settled only to developmental events occurring in the normal course of embryonic development, as both marketed hOP-1 and hBMP-2 have been discontinued for lack of safety, limited effectiveness in clinical contexts, serious collateral effects and regulatory issues with the FDA (5,27,(45)(46)(47)(48)(49).…”

“…The conundrum of regenerative medicine and tissue engineering was, and still is, based on unfulfilled promises of tissue and/or organ regeneration . As we have repeatedly reported, the tremendous advances in molecular, cellular, developmental and surgical biology have not yet met the acid test of regenerative medicine and tissue engineering, i.e., the predictable and routinely successful translation in clinical contexts of the novel tissue engineering procedures so brilliantly tested and developed in preclinical animal models, including non‐human primates .…”