Soo-Woo Kim

Tissue-engineered growth of cartilage: the effect of varying the concentration of chondrocytes seeded onto synthetic polymer matrices

Ninety-six synthetic bioresorbable cell-delivery devices (10 x 10 x 0.5 mm) were seeded, varying the concentrations of living chondrocytes (2, 10, 20, 100 million cells/cc) isolated from shoulders of freshly killed calves and implanted subcutaneously on the dorsum of nude mice after 1 week of in vitro culture. This resulted in the formation of new cartilage in 95.6% of the implants. Twenty-four control implants (0 cells seeded) did not show cartilage formation. During 12 weeks of in vivo implantation, the wet weight and the thickness of the specimens (10, 20, 100 million cells/cc) increased significantly. Histologic analysis revealed cells appearing in their own lacunar structures surrounded by basophilic matrix. The increase in sulfated glycosaminoglycan content indicated the maturation of the extracellular matrix. The ability to manipulate the growth of new cartilage on biocompatible polymer scaffolds by varying the cell density before in vivo implantation will allow engineering to optimize the utilization of chondrocytes in relation to the desired shape, thickness, and quality of the new cartilage.

The acrosomal vesicle of mouse sperm is a calcium store

Subsequent to binding to the zona pellucida, mammalian sperm undergo a regulated sequence of events that ultimately lead to acrosomal exocytosis. Like most regulated exocytotic processes, a rise in intracellular calcium is sufficient to trigger this event although the precise mechanism of how this is achieved is still unclear. Numerous studies on mouse sperm have indicated that a voltage‐operated Ca2+ channel plays some immediate role following sperm binding to the zona pellucida glycoprotein ZP3. However, there is also evidence that the mammalian sperm acrosome contains a high density of IP3 receptors, suggesting that the exocytotic event involves the release of Ca2+ from the acrosome. The release of Ca2+ from the acrosome may directly trigger exocytosis or may activate store‐operated Ca2+ channels in the plasma membrane. To test the hypothesis that the acrosome is an intracellular store we loaded mammalian sperm with the membrane permeant forms of three Ca2+‐sensitive fluorescent indicator dyes: fura‐2, indo‐1, and Calcium Green‐5N. Fluorescence microscopy revealed that the sperm were labeled in all intracellular compartments. When fura‐2 labeled sperm were treated with 150 μM MnCl2 to quench all fluorescence in the cytosol, or when the sperm were labeled with the low affinity dye Calcium Green‐5N, there was a large Ca2+ signal in the acrosome. Consistent with the acrosome serving as an intracellular Ca2+ reservoir, the addition of 20 μM thapsigargin, a potent inhibitor of the smooth endoplasmic reticular Ca2+‐ATPase (SERCA), to populations of capacitated sperm resulted in nearly 100% acrosomal exocytosis within 60 min (τ1/2 ∼ 10 min), in the absence of extracellular Ca2+. Additionally, treatment of sperm with 100 μM thimerosal, an IP3 receptor agonist, also resulted in acrosomal exocytosis. Taken together, these data suggest that the mouse sperm acrosome is a Ca2+ store that regulates its own exocytosis through an IP3 Ca2+ mobilization pathway.

Factors Associated With Dental Implant Survival: A 4-Year Retrospective Analysis

BACKGROUND Dental implants are a predictable treatment option for replacing missing teeth and have strong survival and success outcomes. However, previous research showed a wide array of potential risk factors that may have contributed to dental implant failures. The objectives of this study are to study if implant survival rates were affected by known risk factors and risk indicators that may have contributed to implant failures. The secondary outcome measures were whether the level of expertise of the periodontal residents affected success rates and how the rate of implant success at the Harvard School of Dental Medicine (HSDM) compared to published standards. METHODS A retrospective chart review of patients at the HSDM who had one of two types of rough-surface implants (group A or B) placed by periodontology residents from 2003 to 2006 was performed. Demographic, health, and implant data were collected and analyzed by multimodel analyses to determine failure rates and any factors that may have increased the likelihood of an implant failure. RESULTS The study cohort included 341 dental implants. The odds ratio for an implant failure was most clearly elevated for diabetes (2.59 implant surface group B (7.84), and male groups (4.01). There was no significant difference regarding the resident experience. The success rate for HSDM periodontology residents was 96.48% during the 4-year study period. CONCLUSIONS This study demonstrates that implant success rates at HSDM fell within accepted published standards, confirmed previously identified risk factors for a failure, and potentially suggested that other acknowledged risk factors could be controlled for. Furthermore, the level of experience of the periodontology resident did not have an impact on survival outcomes.

Defining recovery neurobiology of injured spinal cord by synthetic matrix-assisted hMSC implantation.

Significance We developed a platform technology to determine therapeutic mechanisms of human mesenchymal stromal stem cells (hMSCs) in a dorsal root ganglion coculture system and an intraspinal cord implantation model. The unique poly(lactic-co-glycolic) acid scaffolding augments hMSC stemness, engraftment, and function without neural transdifferentiation or mesenchymal lineage development, resulting in robust motosensory improvement, pain and tissue damage mitigation, and myelin preservation in adult rat spinal cord after injury. The scaffolded hMSC-derived neurotrophism, neurogenesis, angiogenesis, antiautoimmunity, and antiinflammation support the propriospinal network, neuromuscular junctions, and serotonergic reticulospinal reinnervation to activate the central pattern generator for restoring hindlimb locomotion. Our findings illuminate “recovery neurobiology”—i.e., the injured spinal cord may deploy polysynaptic neural circuits different from normal adulthood pathways for postinjury improvement. Mesenchymal stromal stem cells (MSCs) isolated from adult tissues offer tangible potential for regenerative medicine, given their feasibility for autologous transplantation. MSC research shows encouraging results in experimental stroke, amyotrophic lateral sclerosis, and neurotrauma models. However, further translational progress has been hampered by poor MSC graft survival, jeopardizing cellular and molecular bases for neural repair in vivo. We have devised an adult human bone marrow MSC (hMSC) delivery formula by investigating molecular events involving hMSCs incorporated in a uniquely designed poly(lactic-co-glycolic) acid scaffold, a clinically safe polymer, following inflammatory exposures in a dorsal root ganglion organotypic coculture system. Also, in rat T9–T10 hemisection spinal cord injury (SCI), we demonstrated that the tailored scaffolding maintained hMSC stemness, engraftment, and led to robust motosensory improvement, neuropathic pain and tissue damage mitigation, and myelin preservation. The scaffolded nontransdifferentiated hMSCs exerted multimodal effects of neurotrophism, angiogenesis, neurogenesis, antiautoimmunity, and antiinflammation. Hindlimb locomotion was restored by reestablished integrity of submidbrain circuits of serotonergic reticulospinal innervation at lumbar levels, the propriospinal projection network, neuromuscular junction, and central pattern generator, providing a platform for investigating molecular events underlying the repair impact of nondifferentiated hMSCs. Our approach enabled investigation of recovery neurobiology components for injured adult mammalian spinal cord that are different from those involved in normal neural function. The uncovered neural circuits and their molecular and cellular targets offer a biological underpinning for development of clinical rehabilitation therapies to treat disabilities and complications of SCI.

Stemness enhancement of human neural stem cells following bone marrow MSC coculture.

Rapid loss of stemness capacity in purified prototype neural stem cells (NSCs) remains a serious challenge to basic and clinical studies aiming to repair the central nervous system. Based on the essential role of mesodermal guidance in the process of neurulation, we hypothesized that coculture of human NSCs (hNSCs) with human bone marrow-derived mesenchymal stromal stem cells (hMSCs) could enhance the stemness of hNSCs through Notch-1 signaling. We have now tested the hypothesis by assessing behaviors of hNSCs and hMSCs under systematically designed coculture conditions relative to monocultures, with or without Notch-1 manipulation in vitro. Our data demonstrate that expression levels of Notch-1 and Hes-1 as determined by immunocytochemistry are significantly higher in hNSCs cocultured with hMSCs than those of controls. Furthermore, coculturing significantly increases immunoreactivity of CD15, a neural stemness marker, but decreases CD24, a marker of neural/neuronal commitment in hNSCs. The effect is independent from the physical status of cell growth since coculture and notch signaling actually promotes hNSC adhesion. Importantly, coculture with hMSCs markedly augments hNSC proliferation rate (e.g., higher yield in G2/M phase subpopulation in a notch-dependent manner detected by flow cytometry) without diminishing their lineage differentiation capabilities. The results suggest that coculture of hNSCs with hMSCs enhances stemness biology of hNSCs partially via activation of Notch-1 signal transduction. Our finding sheds new light on mesoderm-ectoderm cell fate determination via contact-based hMSC-hNSC interactions and provides mechanistic leads for devising effective regimens to sustain and augment stemness of in vitro established hNSC and hMSC lines for basic science, translational and clinical applications.

A Prospective, Randomized Controlled Preclinical Trial to Evaluate Different Formulations of Biphasic Calcium Phosphate in Combination With a Hydroxyapatite Collagen Membrane to Reconstruct Deficient Alveolar Ridges

Many patients and clinicians would prefer a synthetic particulate bone replacement graft, but most available alloplastic biomaterials have limited osteogenic potential. An alloplast with increased regenerative capacity would be advantageous for the treatment of localized alveolar ridge defects. This prospective, randomized controlled preclinical trial utilized 6 female foxhounds to analyze the osteogenic impact of different formulations of biphasic calcium phosphate (BCP) in combination with an hydroxyapatite-collagen membrane and their ability to reconstruct deficient alveolar ridges for future implant placement. The grafted sites were allowed to heal 3 months, and then trephine biopsies were obtained to perform light microscopic and histomorphometric analyses. All treated sites healed well with no early membrane exposure or adverse soft tissue responses during the healing period. The grafted sites exhibited greater radiopacity than the surrounding native bone with BCP particles seen as radiopaque granules. The graft particles appeared to be well-integrated and no areas of loose particles were observed. Histologic evaluation demonstrated BCP particles embedded in woven bone with dense connective tissue/marrow space. New bone growth was observed around the graft particles as well as within the structure of the graft particulate. There was intimate contact between the graft particles and newly formed bone, and graft particles were bridged by the newly formed bone in all biopsies from the tested groups. The present study results support the potential of these BCP graft particulates to stimulate new bone formation. Clinical studies are recommended to confirm these preclinical findings.

The Combination of Purified Recombinant Human Platelet-Derived Growth Factor-BB and Equine Particulate Bone Graft for Periodontal Regeneration

BACKGROUND The objective of this study is to evaluate the potential for periodontal regeneration of a critical-sized defect with the application of recombinant human platelet-derived growth factor (rhPDGF-BB) combined with either a particulate equine or a β-tricalcium phosphate (β-TCP) matrix. METHODS Critical-sized intrabony 2-wall defects were created bilaterally on the distal surface of the second premolar and the mesial surface of the first molar in nine hounds. Twelve defects received rhPDGF-BB/equine treatment, 12 defects received rhPDGF-BB/β-TCP treatment, and the remaining 12 sites served as sham-surgery controls. The animals were sacrificed after a 10-week healing period. RESULTS Clinical healing was uneventful without obvious signs of overt gingival inflammation. Histologic and histomorphometric analyses revealed statistically that there were differences among the three groups in terms of new bone formation (P <0.001). The amount of test material for both rhPDGF-BB/equine and rhPDGF-BB/β-TCP groups was comparable, but the amount of newly formed bone was significantly higher (P <0.01) in favor of the rhPDGF-BB/equine group. The amount of new cementum formed for the rhPDGF-BB/equine group (4.8 ± 1.3 mm) was significantly higher (P =0.001) than the sham-surgery control group (1.7 ± 1.9 mm). CONCLUSION Both rhPDGF-BB/equine and rhPDGF-BB/β-TCP have the potential to support the regeneration of the periodontal attachment apparatus.

A prospective 9-month human clinical evaluation of Laser-Assisted New Attachment Procedure (LANAP) therapy.

This investigation was designed and implemented as a single-center, prospective study to evaluate the clinical response to the Laser-Assisted New Attachment Procedure (LANAP). Eight patients with advanced periodontitis were enrolled and treated with full-mouth LANAP therapy and monitored for 9 months. Fullmouth clinical measurements, including clinical attachment level (CAL), probing depth (PD), and recession, were provided at baseline and after 9 months of healing by a single calibrated examiner, including a total of 930 sites and 444 sites with initial PD equal to or greater than 5 mm. Clinical results for the 930 sites measured pre- and postoperatively revealed that mean PD was reduced from 4.62 ± 2.29 mm to 3.14 ± 1.48 mm after 9 months (P < .05). CAL decreased from 5.58 ± 2.76 mm to 4.66 ± 2.10 mm (P < .05) and recession increased from 0.86 ± 1.31 mm to 1.52 ± 1.62 after 9 months (P < .05). For the subset of 444 sites with initial PD greater than or equal to 5 mm, the PD decreased from 6.50 ± 2.07 mm to 3.92 ± 1.54 mm (P < .05) and CAL decreased from 7.42 ± 2.70 mm to 5.78 ± 2.06 mm (P < .05). As demonstrated by the clinical evaluation, the majority of treated sites demonstrated clinical improvement. LANAP therapy should be further investigated with long-term clinical trials to compare the stability of clinical results with conventional therapy.

A 10-year retrospective radiographic study of implantium dental implants.

A retrospective radiographic study was conducted from five private dental offices on patients requesting dental implant therapy to replace their missing teeth. All implants were placed and restored by early 2000, and patients were encouraged to continue their dental care at the same office. The records were reviewed and analyzed by the clinicians, who had more than 15 years of dental experience at the time of the initial patient treatment. A total of 74 patients with 242 implants were recalled up to 10 years (mean follow-up: 9.21 ± 1.7 years) after loading. There were five implant failures from this radiographic observation period, resulting in a 97.9% dental implant survival rate. The mean crestal bone level change on the mesial aspect was -0.36 ± 1.05 mm, while the mean crestal bone level change on the distal aspect was -0.18 ± 0.96 mm. Thus, the overall mean bone loss was -0.28 ± 0.05 mm. The dental implants, which had a sandblasted, large-grit, acid-etched surface, appeared to achieve successful osseointegration in this long-term observation period, and the implant systems unique design and surface features resulted in a stable osseous crest without bone loss to the first thread.

The Clinical and Histologic Outcome of Dental Implant in Large Ridge Defect Regenerated With Alloplast: A Randomized Controlled Preclinical Trial

A basic tenet of successful osseointegration is that the implant resides in a sufficient quality and quantity of bone to ensure bone contact and thus stabilization. A prospective, randomized controlled preclinical trial was conducted to evaluate the bone-to-implant contact (BIC) when placing implants in bone regenerated by 3 different combinations of biphasic calcium phosphate (BCP). Dental implants were placed into the regenerated ridges of 6 female foxhounds; the ridges were reconstructed with different formulations of BCP in combination with an hydroxyapatite collagen membrane. They were retrieved after 3 months to perform light microscopic and histomorphometric analyses. Implants in each group appeared to be stable and osseointegrated. Light microscopic evaluation revealed tight contacts between the implant threads with the surrounding bone for all 4 groups. The mean BIC ranged from 64.7% to 73.7%. This preclinical trial provided clinical and histologic evidence to support the efficacy of all 3 formulations of BCP to treat large alveolar ridge defects to receive osseointegrated dental implants.