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Prolonged In Vitro Expansion Shapes the Neuro-Supportive Potential of Jaw Periosteum Secretomes: Implications for Secretome Product Quality.

Nerve injuries are frequent complications of complex oral and maxillofacial surgical procedures, particularly following extensive tumor resections. Secretome-based, cell-free therapies derived from mesenchymal stromal cells have emerged as promising regenerative approaches; however, robust manufacturing requires the identification of critical quality attributes (CQAs) that ensure product potency and consistency. The influence of replicative senescence during in vitro expansion on the quality of jaw periosteum-derived mesenchymal stromal cell (JPC) secretomes has not yet been established. This study investigated whether the expansion state of JPCs affects the composition and neuro-supportive potency of their secretomes. Secretomes from four independent JPC donors were collected separately at early and late passages, pooled within each passage-specific preparation, and applied to human induced pluripotent stem cell-derived neurons. Neuronal survival, neurite outgrowth, and neuronal marker expression were assessed as functional readouts. Secretome composition was characterized by quantitative proteomics and enzyme-linked immunosorbent assay (ELISA) of selected senescence-associated secretory phenotype (SASP) factors. Secretomes derived from early-passage JPCs significantly enhanced neuronal survival and neurite outgrowth, whereas late-passage secretomes displayed reduced neuro-supportive activity. Proteomic profiling identified a pronounced shift toward inflammatory and stress-associated signaling, whereas performed ELISAs confirmed senescence-associated remodeling of the secretome, including increased abundance of SASP-associated factors in late-passage preparations. These findings demonstrate that prolonged in vitro expansion profoundly influences both the composition and biological potency of JPC-derived secretomes. Collectively, this study identifies the passage-associated senescence-like phenotype of JPCs as a key determinant of secretome quality and supports its consideration as a critical quality attribute for the manufacturing and standardization of JPC-derived secretome products. Monitoring and controlling the expansion state of JPCs may therefore be essential to ensure the consistency, potency, and clinical translation of secretome-based regenerative therapies.

Humans

[Histological studies on the healing of mandibular osteotomies in the dog following stable osteosynthesis with the cold polymerizing polymethylmethacrylate "Kallokryl K"].

The disadvantages of methods of accomplishing stable osteosynthesis of mandibular fractures are described and the possibilities of jacketing the fragments with Kallokryl K plastic material, which is a cold-polymerizing polymethyl methacrylate, pointed out. After six months from the surgical operation, the healing under Kallokryl K was clinically and histologically evaluated on canine mandibles subjected to osteotomy and with the periosteum removed from the lower jaw bone. The reaction of both soft tissue and bone to the plastic material attached thereto was also evaluated. The results are discussed in the light of what is presently known about primary and secondary bone healing.

Animals

Periostitis of the mandible. A case report and review of the literature.

Periostitis of the mandible has not been previously discussed in the otolaryngologic literature and only infrequently in the dental literature. It is characterized by a hard, painless, fixed mass overlying the mandible in a child or young adult. The etiology is usually an endosteal infection in the molar region with periosteal inflammation. Reactive new bone is deposited beneath the elevated periosteum, external to the outer cortex of the jaw, and separated from it by a layer of suppurative fluid. Treatment consists of removal of involved teeth and infected bone, drainage of the fluid pocked and appropriate antibiotics. Patients are followed with repeat X-rays and erythrocyte sedimentation rates. A case is presented and the world literature is reviewed. The otolaryngologist should be aware of this entity and include it in the differential diagnosis of any fixed jaw mass to avoid mistaking it for a more malignant process with unfortunate consequences.

Adolescent

In vitro studies on skeletogenic potential of membrane bone periosteal cells.

In the avian embryo ectomesenchyme cells, derived from the mesencephalic level of the cranial neural crest, migrate into the presumptive maxillary region and subsequently differentiate into the membrane bones and associated secondary cartilage of the upper jaw skeleton. The cartilage arises secondarily within the periosteum at points of articulation between membrane bones and provides an embryonic articulating surface. The stimulus for the differentiation of secondary cartilage is believed to be intermittent pressure and shear created at the developing embryonic movement. The development of one such system--the quadratojugal, has been analysed using organ and explant culture techniques and studied with particular reference to the differentiation of periosteal cells into secondary cartilage. A number of conclusions were reached. (1) Normally only cells at discrete loci express a chondrogenic potential in vivo: the periosteal cells at these sites of future articulation become committed to chondrogenesis during stage 35, more than 24 h before cartilage is identifiable in vivo. (2) However, cells with a 'latent' chondrogenic potential are widespread in membrane bone periosteum and occur over most, if not all, of the surface area of the bone. This potential is expressed in the 'permissive' environment created by submersion of the tissue in explant culture or in submerged organ culture. (3) This chondrogenic potential exists long before the time at which commitment of cartilage-forming cells occurs and even presumptive maxillary ectomesenchyme at stage 29 has a limited ability to form cartilage in vitro. It is suggested that spatial position is a principal factor controlling the differentiation of secondary cartilage. Ectomesenchyme cells with the potential to form secondary cartilage are widespread but it is only those cells whose migration from the neural crest positions them and their progeny at the site of a presumptive joint which subsequently express this potential. This epigenetic interpretation is discussed in the general context of development mechanisms underlying the spatial and temporal patterns in which neural crest-derived cells differentiate to produce bone and cartilage during the formation of the head skeleton.

Animals

Experimental subperiosteal implantation of porous Al2O3 ceramic for mandibular ridge augmentation.

Porous Al2O3 ceramic material with pore size of 100-750 micrometers was used for implants placed subperiosteally as alveolar ridge augmentation onlays in six Cercopithecus monkeys. Clinical and radiographic examination during the postoperative period revealed rapid healing and anchorage of the implant in five of the animals. One implant was lost. The implants were retrieved after 4, 10, 11 and 12 months' implantation time. Histologic and microradiographic examination demonstrated fibrous and bone tissue invasion into the pore system at the base of the subperiosteal implant, with good anchorage of the ceramic material to the jaw. Only fibrous tissue was found to project into the ceramic openings on the implant sides facing the periosteum. Labeling of the mineralizing tissue was attempted using intravenously administered tetracycline (Reverin) and calcein (Calcein) at varying intervals prior to sacrifice, but the labeling results were not impressive.

Aluminum Oxide

Ultrastructure of tissue in intrabony and subperiosteal porous ceramic implants.

Porous ceramic (Al2O3) implants with pore size of 100--750 micron were placed intrabony in the maxilla and subperiosteally in the mandible of three adult monkeys. After observation periods of 10, 11 and 12 months, the animals were perfused with 1.7% glutaraldehyde and the jaws processed for electron microscopy. Tissue ingrowth into the pores was observed in all the specimens. The implant pores facing the host bone contained bone tissue to a depth of approximately 1 mm, while only unmineralized tissue was found in the implant part adjacent to the periosteum. The unmineralized tissue was of a fibrous nature with collagen fibers arranged in a lamellar fashion. Cells, often with extremely long and slender cellular processes, were seen between the lamellae. The cells had the cytoplasmic features typical of protein-producing cells. The mineralized tissue exhibited ordinary bone tissue structure with plate-like crystallites in a collagenous matrix. Some lacunae and canaliculi with osteocytes were observed, though they were scarce.

Animals

A modified pre-auricular approach to the temporomandibular joint and malar arch.

In an attempt to improve visibility and safety in the surgical approach to the malar arch and jaw joint, anatomical dissections of 56 facial halves were undertaken. Observations are made on the relationship of the facial nerve bifurcation and its temporal branch to bony landmarks. Attention is drawn to the dangerous area of fusion of the superficial fascia, the superficial layer of temporal fascia and the periosteum of the malar arch. The safety of approaching the malar arch through the pocket formed by the splitting of the lower part of the temporal fascia is emphasised. Minor modifications to the established pre-auricular approach were made and applied successfully to six sides in five patients.

Facial Nerve

Influences of different surgical procedures on growth of dentomaxillary complex in dogs with artificially created cleft palate.

Possible detrimental effects of palate repair on dentomaxillary growth have been reported before. In our study, two principally different surgical techniques were compared. Dogs with artificially created clefts of the palate were subjected to either a periosteal flap (Langenbeck) or to a bone flap (Dieffenbach) closure. Skull roentgenograms and upper jaw impressions were used to study the possible effects on growth. Definite abnormalities were found following both procedures; there are strong indications that the bone flap is the more detrimental one.

Animals

Functional significance of the mandibular symphysis.

The purpose of this investigation was to relate the morphology of connective tissues in the mandibular symphysis to the behavioral and experimental evidence for mobility and mechanical stress at the symphysis. The anatomy of the symphysis was examined histologically in 6 mammalian orders encompassing 22 species. Behavioral and experimental evidence of stress during the power stroke of the chewing cycle correspond with stresses at the symphysis implied by the location and orientation of symphyseal connective tissues. These stresses are: (1) dorsoventral shear of the symphysis due to the transfer of force from balancing to chewing sides, (2) bending of the symphysis causing tension along the inferior and compression along superior borders due to torsion on the dentaries from the jaw closing muscles, and (3) antero-posterior shear of the symphysis due to an anteriorly directed stress on the chewing side. Interspecific comparisons suggest that leaf eaters can resist greater dorsoventral shear than fruit or insect eaters, but no correlations exist between diet and bending or antero-posterior shear. This suggests that chewing leaves requires larger biting forces.

Animals