Connexin 43 regulates the expression of wound healing-related genes in human gingival and skin fibroblasts.

Fibroblasts are the most abundant connective tissue cells and play an important role in wound healing. It is possible that faster and scarless wound healing in oral mucosal gingiva relative to skin may relate to the distinct phenotype of the fibroblasts residing in these tissues. Connexin 43 (Cx43) is the most ubiquitous Cx in skin (SFBLs) and gingival fibroblasts (GFBLs), and assembles into hemichannels (HCs) and gap junctions (GJs) on the cell membrane. We hypothesized that SFBLs and GFBLs display distinct expression or function of Cx43, and that this may partly underlie the different wound healing outcomes in skin and gingiva. Here we show that Cx43 distinctly formed Cx43 GJs and HCs in human skin and gingiva in vivo. However, in SFBLs, in contrast to GFBLs, only a small proportion of total Cx43 assembled into HC plaques. Using an in vivo-like 3D culture model, we further show that the GJ, HC, and channel-independent functions of Cx43 distinctly regulated wound healing-related gene expression in GFBLs and SFBLs. Therefore, the distinct wound healing outcomes in skin and gingiva may partly relate to the inherently different assembly and function of Cx43 in the resident fibroblasts.

Elevated CD26 Expression by Skin Fibroblasts Distinguishes a Profibrotic Phenotype Involved in Scar Formation Compared to Gingival Fibroblasts.

Compared to skin, wound healing in oral mucosa is faster and produces less scarring, but the mechanisms involved are incompletely understood. Studies in mice have linked high expression of CD26 to a profibrotic fibroblast phenotype, but this has not been tested in models more relevant for humans. We hypothesized that CD26 is highly expressed by human skin fibroblasts (SFBLs), and this associates with a profibrotic phenotype distinct from gingival fibroblasts (GFBLs). We compared CD26 expression in human gingiva and skin and in gingival and hypertrophic-like scar-forming skin wound healing in a pig model, and used three-dimensional cultures of human GFBLs and SFBLs. In both humans and pigs, nonwounded skin contained abundantly CD26-positive fibroblasts, whereas in gingiva they were rare. During skin wound healing, CD26-positive cells accumulated over time and persisted in forming hypertrophic-like scars, whereas few CD26-positive cells were present in the regenerated gingival wounds. Cultured human SFBLs displayed significantly higher levels of CD26 than GFBLs. This was associated with an increased expression of profibrotic genes and transforming growth factor-ß signaling in SFBLs. The profibrotic phenotype of SFBLs partially depended on expression of CD26, but was independent of its catalytic activity. Thus, a CD26-positive fibroblast population that is abundant in human skin but not in gingiva may drive the profibrotic response leading to excessive scarring.

Sterilization of Ceramic Sharpening Stones.

Traditionally, periodontal hand instruments are honed or sharpened during patient care as they dull easily during contact with enamel, calculus and cementum. This approach is taught in dental and hygiene schools around the world and remains the standard of care. Recently, some professional organizations have questioned whether this practice should be abandoned because of safety issues. Questions have been raised whether sharpening stones can be properly sterilized and whether the sharpening of contaminated instruments poses a health hazard for the provider. Using bacteria culture techniques and scanning electron microscopy, we tested whether contaminated ceramic sharpening stones can be sterilized. Our results demonstrate that the stones were sterile after being subjected to the manufacturer's sterilization protocol. In addition, over the last year, no incidents related to periodontal instrument sharpening have been reported among nearly 400 students at the faculty of dentistry, University of British Columbia, where chair-side sharpening is taught. Therefore, we conclude that ceramic sharpening stones can be sterilized using normal office protocols and that chair-side sharpening adds little risk beyond routine handling of operatory or periodontal instruments during patient care when proper protocols are followed.

Critical role for αvß6 integrin in enamel biomineralization.

Tooth enamel has the highest degree of biomineralization of all vertebrate hard tissues. During the secretory stage of enamel formation, ameloblasts deposit an extracellular matrix that is in direct contact with the ameloblast plasma membrane. Although it is known that integrins mediate cell-matrix adhesion and regulate cell signaling in most cell types, the receptors that regulate ameloblast adhesion and matrix production are not well characterized. We hypothesized that αvß6 integrin is expressed in ameloblasts where it regulates biomineralization of enamel. Human and mouse ameloblasts were found to express both ß6 integrin mRNA and protein. The maxillary incisors of Itgb6(-/-) mice lacked yellow pigment and their mandibular incisors appeared chalky and rounded. Molars of Itgb6(-/-) mice showed signs of reduced mineralization and severe attrition. The mineral-to-protein ratio in the incisors was significantly reduced in Itgb6(-/-) enamel, mimicking hypomineralized amelogenesis imperfecta. Interestingly, amelogenin-rich extracellular matrix abnormally accumulated between the ameloblast layer of Itgb6(-/-) mouse incisors and the forming enamel surface, and also between ameloblasts. This accumulation was related to increased synthesis of amelogenin, rather than to reduced removal of the matrix proteins. This was confirmed in cultured ameloblast-like cells, in which αvß6 integrin was not an endocytosis receptor for amelogenins, although it participated in cell adhesion on this matrix indirectly via endogenously produced matrix proteins. In summary, integrin αvß6 is expressed by ameloblasts and it plays a crucial role in regulating amelogenin deposition and/or turnover and subsequent enamel biomineralization.

Intra-operative application of chlorhexidine gel reduces bacterial counts in internal implant cavity.

A prospective clinical trial was conducted to assess the bacterial-inhibitory potential of 1% chlorhexidine (CHX) gel in the internal cavity of implant screw holes, when utilized at the time of implant placement. A total of 40 Straumann (S) and Nobel Biocare (N) implants were divided into test (ST or NT; implant + CHX gel) and control (SC or NC; implant only) groups. Total numbers of colony-forming units (CFUs ml(-1) ) were assessed at a minimum of 3 months postsurgery by aerobic and anaerobic culture. A set of specimens was stained with Gram stain. The mean sample-collection time was 110 d for the test population and 98 d for the controls. The use of 1% CHX gel significantly reduced bacterial counts in both the ST and NT samples by over three logs compared with controls. No statistical differences in the numbers of CFUs ml(-1) were evident between aerobic and anaerobic cultures. Differences in the numbers of CFUs ml(-1) between ST and NT groups were not statistically significant. Microscopic analysis showed mainly Gram-positive coccoid species in most samples.

Scanning electron microscopic analysis of adherent bacterial biofilms associated with peri-implantitis.

OBJECTIVES: Peri-implantitis (PI) is caused by bacteria in the peri-implant space but the consensus on microbial profile is still lacking. Current microbial sampling of PI lesions has largely focused on analyzing bacterial species that have been shed from the implant surface and captured in the pocket fluid. The purpose of the present study was to investigate the morphotypes of bacteria in biofilm covering the implant threads and explore whether certain morphotypes were associated with PI. METHODS: Fourteen failed implants were removed and instantly processed for scanning electron microscope analysis. The implants were imaged at three equally divided sub-crestal levels of the exposed area. Bacterial morphotypes were identified and quantified by three examiners. Mobility and years in function were correlated to the presence of different morphotypes. RESULTS: The implants demonstrated the presence of variable bacterial morphotypes that did not correlate to disease progression in our study. Some implants were dominated by filaments and others showed the presence of combinations of cocci/rods or spirilles/spirochetes. In general, all implants showed variable morphologic biofilm composition. However, individual implants tended to have similar composition throughout the entire implant. Rods and filaments were dominant morphotypes throughout the surfaces and cocci showed increased presence toward the apical third. There were some differences in the biofilm morphology with mobility and time in function. CONCLUSIONS: The profiles of bacterial biofilm morphotypes in failing implants with similar clinical presentations were highly variable. While there were significant differences between implants, similar morphotypes in individual implants were often found throughout the entire surface.

Exploring scarless healing of oral soft tissues.

Our research group is comparing clinical, histological and molecular healing profiles of oral and skin wounds using human and pig models. The goal is to determine the molecular cues that lead to scarless healing in the oral mucosa and use that information to develop scar prevention therapies for skin and prevent aberrant wound healing in the oral cavity. Wound healing in human and pig palatal mucosa is almost identical, and scar formation is reduced in oral wounds compared with skin. The striking difference between these tissues is transient and rapidly resolving inflammation in oral wounds compared with long-lasting inflammation in the skin wounds. Currently, we are looking at wound transcriptomes (genes differentially regulated) and proteomes (a set of proteins) to investigate how these wound healing responses in skin and oral mucosa are regulated at the molecular level.

Expression of integrin alphavbeta6 and TGF-beta in scarless vs scar-forming wound healing.

Oral mucosal wounds heal with reduced scar formation compared with skin. The epithelial integrin alphavbeta6 is induced during wound healing, and it can activate fibrogenic transforming growth factor beta1 (TGF-beta1) and anti-fibrogenic TGF-beta3 that play key roles in scar formation. In this study, expression of beta6 integrin and members of the TGF-beta pathway were studied in experimental wounds of human gingiva and both gingiva and skin of red Duroc pigs using real-time PCR, gene microarrays, and immunostaining. Similar to human wounds, the expression of beta6 integrin was induced in the pig wounds 7 days after wounding and remained upregulated >49 days. The alphavbeta6 integrin was colocalized with both TGF-beta isoforms in the wound epithelium. Significantly higher expression levels of beta6 integrin and TGF-beta1 were observed in the pig gingival wounds compared with skin. Early gingival wounds also expressed higher levels of TGF-beta3 compared with skin. The spatio-temporal colocalization of alphavbeta6 integrin with TGF-beta1 and TGF-beta3 in the wound epithelium suggests that alphavbeta6 integrin may activate both isoforms during wound healing. Prolonged expression of alphavbeta6 integrin along with TGF-beta3 in the gingival wound epithelium may be important in protection of gingiva from scar formation.

Wound healing in oral mucosa results in reduced scar formation as compared with skin: evidence from the red Duroc pig model and humans.

Scar formation is a common, unwanted result of wound healing in skin, but the mechanisms that regulate it are still largely unknown. Interestingly, wound healing in the oral mucosa proceeds faster than in skin and clinical observations have suggested that mucosal wounds rarely scar. To test this concept, we created identical experimental wounds in the oral mucosa and skin in red Duroc pigs and compared wound healing and scar development over time. We also compared the pig oral mucosal wound healing to similar experimental wounds created in human subjects. The findings showed significantly reduced scar formation at both clinical and histological level in the pig oral mucosa as compared with skin 49 days after wounding. Additionally, the skin scars contained a significantly increased number of type I procollagen immunopositive cells and an increased fibronectin content, while the oral mucosal wounds demonstrated a prolonged accumulation of tenascin-C. Furthermore, the pig oral mucosal wounds showed similar molecular composition and clinical and histological scar scores to human oral mucosal wounds. Thus, the reduced scar formation in the pig oral mucosa provides a model to study the biological processes that regulate scarless wound healing to find novel approaches to prevent scar formation in skin.

Localization and potential function of kindlin-1 in periodontal tissues.

Kindlin-1 is an intracellular focal adhesion protein that regulates the actin cytoskeleton. Patients suffering from Kindler syndrome have a homologous mutation of the kindlin-1 gene and develop skin blisters, periodontal disease, and intestinal complications because of deficient adhesion of the basal epithelial cells. We investigated kindlin-1 localization in periodontal tissue and its functions in cultured keratinocytes and showed that kindlin-1 co-localizes with migfilin and paxillin in the basal epithelial cells of oral mucosa and in cultured keratinocytes. The kindlin-1-deficient oral mucosal tissue from a patient with Kindler syndrome showed a complete lack of paxillin and reduced migfilin immunostaining in the basal keratinocytes. Co-immunoprecipitation showed that migfilin directly interacted with kindlin-1. RNA interference-induced kindlin-1 deficiency in keratinocytes led to an altered distribution of migfilin-containing focal adhesions, reduced cell spreading, decreased cell proliferation, and decelerated cell migration. Disruption of microtubules in the kindlin-1-deficient cells further reduced cell spreading, suggesting that microtubules can partially compensate for kindlin-1 deficiency. Kindlin-1 supported mature cell-extracellular matrix adhesions of keratinocytes, as downregulation of kindlin-1 expression significantly reduced the cell-adhesion strength. In summary, kindlin-1 interacts with migfilin and plays a crucial role in actin-dependent keratinocyte cell adhesion essential for epidermal and periodontal health.

Initial torque stability of a new bone condensing dental implant. A cohort study of 140 consecutively placed implants.

The aim of this paper was to determine the torque resistance of this new implant during placement in different types of bone, immediate placement into sockets, and in grafted bone. The torque at time of placement serves as an indication of initial stability, which is accepted as an important factor for implant osseointegration and immediate loading. Within a 13-month period, 140 NobelActive implants in 84 consecutive patients were placed into types I-IV bone in fresh sockets, and into grafted bone (both in maxillary sinuses and on the facial alveolar surfaces where bone had been lost). The final torque was measured with a manual torque control wrench as manufactured by Nobel Biocare for clinical use with this type of implant. One hundred forty implants with 3.5 to 5 mm diameters and 10 to 15 mm lengths were placed in different types of bone, either as delayed or immediate implants into fresh extraction sockets. These implants demonstrated a mean torque stability value of 50.8 Ncm. The average insertion torque for delayed implants was 49.7 Ncm. For immediate implants the average torque was 52.6 Ncm. Placement into soft bone was also favorable at an average of 47.9 Ncm. Typical straight walled and tapered implants generally exhibit 10 to 35 Ncm insertion torques. The NobelActive implant consistently reaches higher torque levels. This may indicate they are more favorably suited to early provisionalization and loading. Soft bone (type IV) did not seem to decrease significantly the torque of insertion of these implants. Further longer term studies are needed to investigate whether this indeed makes these implants more suited for early provisionalization and loading than traditional root form. Long term studies are also needed to investigate maintenance of bone levels surrounding these implants.

Clinical evaluation of the NobelActive implant system: a case series of 107 consecutively placed implants and a review of the implant features.

The purpose of this paper is to (1) introduce the features of this new implant, (2) investigate the clinical benefits as advertised by the manufacturer in comparison with traditional root form implants, and (3) provide guidelines for its use. One hundred seven NobelActive implants were placed in 67 consecutive patients with type I-IV bone within 8 months. Cases also include implants placed in sinus grafts, ridges with insufficient thickness and facial bone loss and were placed with delayed and immediate loading. Parameters were assessed to determine whether we could confirm the manufacturer's statements on this implant system. Results obtained with 107 implants of 3.5, 4.3, and 5 mm diameters with 10- to 15-mm lengths placed in different types of bone with delayed and immediate loading demonstrated a final insertion torque from 15 to 70 Ncm. All types of bone allowed "redirection" of the implant but were limited in the bone with higher density. According to the manufacturer, this new design of the NobelActive implant has high initial stability, bone condensing properties, redirecting capability, built-in platform shifting, and dual-function prosthetic connections. After investigating these 5 statements within the limits of our study, we were able to confirm these claims, but with some recommendations for the clinical use and placement of these implants.

Kindler syndrome and periodontal disease: review of the literature and a 12-year follow-up case.

BACKGROUND: The association of aggressive periodontitis with Kindler syndrome was based on a single case in 1996 and later confirmed with a larger population. Since then, significant research has greatly increased our understanding of the molecular pathology of this disorder. We review recent advances in the molecular mechanisms of the syndrome and present a maintenance case report of a patient who has been followed in our clinic. METHODS: A female patient who was diagnosed with Kindler syndrome and aggressive periodontitis at the age of 16 years has been followed and treated in our clinic for 12 years. Her main treatment has been maintenance therapy following her initial treatment and restorative work previously documented. Gingival biopsies obtained during the recent extraction of hopeless maxillary molars were used for histologic assessment of gingival tissue attachment apparatus and to isolate gingival fibroblasts. Reverse transcription-polymerase chain reaction (RT-PCR) was performed using these cells to confirm the lack of expression of kindlin-1. RESULTS: RT-PCR showed the total loss of kindlin-1 mRNA in cultured gingival fibroblasts, supporting the clinical diagnosis of Kindler syndrome. Tissue biopsies revealed atypical pocket epithelium. Maintenance therapy has been moderately successful. Teeth that were recently lost had a poor prognosis at the initial assessment. The patient's gingiva and oral mucosa continue to be fragile with episodes of sloughing and inflammation. CONCLUSIONS: Periodontitis in Kindler syndrome responds to maintenance therapy, but the gingiva and oral mucosa continue to display an abnormal appearance with white patches. Histologic findings suggest that the junctional epithelium in Kindler syndrome may be abnormal and could explain why these patients have periodontal disease. Attachment loss progressed around teeth with an initial guarded or poor prognosis. Teeth that started with a good or fair prognosis continue to have a fair prognosis. Limited dental implant treatment is being considered.

Distinctive molecular composition of human gingival interdental papilla.

BACKGROUND: Gingiva is composed of attached and marginal (free) gingiva and interdental papilla. Increasing esthetic demands in dentistry have created a need to restore all parts of the gingiva. However, the interdental papilla has limited regeneration potential compared to other parts of the gingiva. It also is more susceptible to gingival overgrowth, suggesting that it has distinct cellular and molecular properties from other parts of the gingiva. Very little is known about the possible differences in the molecular composition of different parts of the gingiva. METHODS: We compared the expression of a set of key molecules in interdental papilla and marginal gingiva from seven healthy subjects by immunohistochemical staining. RESULTS: In the interdental papilla, immunoreactivity for integrin alphavbeta6 and cytokeratin 19 in the oral epithelium was significantly higher than in marginal gingiva. Expression of type I procollagen, extra domain A (EDA) and extra domain B (EDB) fibronectin isoforms, tenascin-C, transforming growth factor-beta (TGF-beta), connective tissue growth factor (CTGF), and the signaling molecule son-of-sevenless (SOS)-1 also were increased in the interdental papilla. The expression of small leucine-rich proteoglycans decorin, biglycan, fibromodulin, and lumican in the interdental papilla was partially different from the marginal gingiva. CONCLUSIONS: Molecular composition of the interdental papilla is distinct from marginal gingiva. Increased expression of molecules normally induced in wound healing (alphavbeta6 integrin, fibronectin-EDB and -EDA, tenascin-C, type I procollagen, TGF-beta, CTGF, and SOS-1) suggests that the cells in the interdental papilla are in an activated state and/or inherently display a specific phenotype resembling wound healing.

Tensile properties of embryonic epithelia measured using a novel instrument.

We present the first measurements of the tensile properties of embryonic epithelia, data that are crucial to understanding the mechanics of morphogenetic movements. Fine wires were glued to the surface of an intact, live embryo using cyanoacrylate glue, after which the epithelium between the wires was separated from the remainder of the embryo by microsurgery. The wires were then separated from each other in 0.1 microm steps under computer control in order to elongate the tissue at a constant true strain rate. Force was determined from the degree of bending in the wires, and a real-time, image-based feedback system corrected for reductions in elongation that would otherwise have been caused by wire flexure. The instrument was used to determine the tensile properties of epidermis and neuroepithelia from early-stage embryos of the axolotl (Ambystoma mexicanum), a type of amphibian. Monolayer specimens as small as 300 by 500 microm were elongated at physiological strain rates of 5-30% per hour, and the effects of developmental stage, epithelium type, specimen origin, direction of elongation and strain rate were investigated. True strains as high as 50% were observed before tearing began and equivalent moduli for the initial, linear portion of the load resultant versus strain curves ranged from 1 x 10(-3) to 8 x 10(-3) N/m.

Clinical and microbiologic study of periodontitis associated with Kindler syndrome.

BACKGROUND: Little is known about the onset and prevalence of periodontal disease in patients with the rare Kindler syndrome, a genodermatological disorder. This study investigated the level of clinical periodontal attachment in relation to age and presence of putative periodontopathogenic bacteria in individuals with Kindler syndrome. METHODS: Eighteen individuals diagnosed with Kindler syndrome and 13 control subjects, aged 4 to 37 years, from rural Panama received a limited clinical periodontal examination. Subgingival samples were collected for identification of putative periodontal pathogens by polymerase chain reaction. RESULTS: Mild to severe gingivitis was a common finding in all adults of the study population. Seventy-two percent (13/18) of the Kindler patients and 46% (6/13) of the control subjects showed mild to severe periodontal disease (P = 0.001, chi-square test). The onset of periodontitis was earlier and the progression occurred at a faster rate in the Kindler group. There was a strong correlation (r = 0.83) between the level of attachment loss and age in the Kindler group and a weaker correlation (r = 0.66) in the control group. The appearance of gingival tissues suggested atypical periodontitis with spontaneous bleeding and fragile, often desquamative, gingiva. In periodontitis patients, Porphyromonas gingivallis and Diallster pneumosintes tended to occur more frequently in control individuals compared to those with Kindler syndrome. CONCLUSIONS: In the Kindler group, periodontitis had an onset in early teenage years and progressed more rapidly compared to non-Kindler individuals of the same geographic and ethnic group. Clinical and microbiological findings suggest atypical periodontitis in Kindler patients. We propose to include Kindler syndrome in the category of medical disorders predisposing to destructive periodontal disease.

Intentional angulation of an implant to avoid a pneumatized maxillary sinus: a case report.

This case report describes placement of an implant in the posterior maxilla so as to avoid a pneumatized sinus and also to avoid the need for a sinus lift procedure. An 81-year-old woman presented with an edentulous span in the upper right posterior maxilla. She had been missing teeth in this area for many years, and there was a combination of resorption of the alveolar ridge and pneumatization of the maxillary sinus. Eleven years previously, implants had been placed anterior to this region, but the patient was told that implants could not be placed posteriorly unless a sinus lift was done. At the time of the current presentation she was still unwilling to undergo a sinus lift procedure but wanted to know if implants could be placed in the posterior right maxilla. A tomogram obtained with a radiographic stent in place indicated that there was insufficient bone height to allow placement of implants at the usual angulation without a sinus lift. Therefore, to avoid the need for a sinus lift, 2 implants were placed with palatal angulation as guided by a tomographically determined surgical stent. The treatment planning and surgical and restorative techniques are reviewed here. A postoperative tomogram was obtained to determine the final position of the implants. The outcome has been favourable for the patient and the clinicians. In situations where there is sufficient palatal bone medial to the maxillary sinus, placing implants at an angle may prevent the need for a sinus lift procedure, assuming that proper development of an occlusal restorative scheme is possible.

Mechanical effects of cell anisotropy on epithelia.

Theoretical, numerical and experimental methods are used to develop a comprehensive understanding of how cell shape affects the mechanical characteristics of two-dimensional aggregates such as epithelia. This is an important step in relating the mechanical properties of tissues to those of the cells of which they are composed. Statistical mechanics is used to derive formulas for the in-plane stresses generated by tensions gamma along cell-cell interfaces in sheets with anisotropic cellular fabric characterized by average cell aspect ratio kappa. These formulas are then used to investigate self-deformation (strain relaxation) of an anisotropic sheet composed of cells of thickness h and having effective viscosity mu. Finite element simulations of epithelia and of isolated cells and novel relaxation studies of specimens of embryonic epithelia reported herein are consistent with the predictions of the theory. In all cases, geometric factors cause the relaxation responses to be more complex than a single decaying exponential.

Scarless healing of oral mucosa is characterized by faster resolution of inflammation and control of myofibroblast action compared to skin wounds in the red Duroc pig model.

BACKGROUND: Scar formation following skin trauma can have devastating consequences causing physiological and psychosocial concerns. Currently, there are no accepted predictable treatments to prevent scarring which emphasizes a need for a better understanding of the wound healing and scar formation process. OBJECTIVES: Previously it was shown that healing of small experimental wounds in the oral mucosa of red Duroc pigs results in significantly reduced scar formation as compared with equivalent full-thickness skin wounds. In the present study, scar formation was assessed in 17 times larger experimental wounds in both oral mucosa and skin of the red Duroc pigs. METHODS: Equivalent experimental wounds were created in the oral mucosa and dorsal skin of red Duroc pigs, and scar formation, localization and abundance of key wound healing cells, transforming growth factor-beta (TGF-beta) and phosphorylated Smad3 (pSmad3) were assessed. RESULTS: Oral mucosal wounds displayed significantly less clinical and histological scar formation than did the corresponding skin wounds. The number of macrophages, mast cells, TGF-beta and pSmad3 immunopositive cells was significantly reduced in the oral mucosal wounds as compared with skin wounds during the maturation stage of the healing process. Although the number of myofibroblasts was significantly elevated, the oral mucosal wounds showed significantly less contraction than did the skin wounds over time. CONCLUSIONS: Earlier resolution of the inflammatory reaction and reduced wound contraction may promote scarless oral mucosal wound healing. In addition, scar formation likely depends not only on the number of myofibroblasts but also on the extracellular environment which regulates their function.

Multiview robotic microscope reveals the in-plane kinematics of amphibian neurulation.

A new robotic microscope system, called the Frogatron 3000, was developed to collect time-lapse images from arbitrary viewing angles over the surface of live embryos. Embryos are mounted at the center of a horizontal, fluid-filled, cylindrical glass chamber around which a camera with special optics traverses. To hold them at the center of the chamber and revolve them about a vertical axis, the embryos are placed on the end of a small vertical glass tube that is rotated under computer control. To demonstrate operation of the system, it was used to capture time-lapse images of developing axolotl (amphibian) embryos from 63 viewing angles during the process of neurulation and the in-plane kinematics of the epithelia visible at the center of each view was calculated. The motions of points on the surface of the embryo were determined by digital tracking of their natural surface texture, and a least-squares algorithm was developed to calculate the deformation-rate tensor from the motions of these surface points. Principal strain rates and directions were extracted from this tensor using decomposition and eigenvector techniques. The highest observed principal true strain rate was 28 +/- 5% per hour, along the midline of the neural plate during developmental stage 14, while the greatest contractile true strain rate was--35 +/- 5% per hour, normal to the embryo midline during stage 15.