C. Ward Kischer

Perivascular myofibroblasts and microvascular occlusion in hypertrophic scars and keloids

Microvessels in normal skin, granulation tissue, hypertrophic scar, keloid, and mature scar from human subjects were studied by transmission electron microscopy. Comparative observations suggested that most microvessels in hypertrophic scar and keloid are occluded or partially occluded, apparently owing to an excess of endothelial cells. Endothelial cell contraction was also supported by the observations, and perivascular satellite cells (pericytes), some of which were identified as myofibroblasts, were observed in hypertrophic scars and keloids. Among findings from statistical analyses were that 1) the patency of microvessels in hypertrophic scar and granulation tissue is similar, as is that of microvessels in keloid and mature scar, but the patency of all these microvessels is significantly less than that of microvessels in normal skin, and 2) endothelial cell density is greater in nonpatent vessels than in patent vessels. The observed extent of microvascular occlusion supports a previously published theory that hypoxia is involved in the generation of hypertrophic scar.

Fibronectin (FN) in hypertrophic scars and keloids

SummaryFibronectin (FN) distribution was compared among samples of normal human dermis, hypertrophic scar, keloid, and granulation tissues from deep injuries. Localization was established by use of fibronectin antibodies and the indirect immunofluorescence method. Fresh-frozen tissue was sectioned on a cryostat and examined by epifluorescence. Hypertrophic scar and keloid demonstrated heavy deposition of FN, which conformed to the nodular characteristics of the lesions. Intense localization occurred in granulation tissue over fibroblasts which were stellate and vesiculated, and over small blood vessels. FN-staining was weak in areas over fibroblasts which were more rounded and nonvesiculated. Staining for FN was also minimal over the collagen in normal dermis and the deeper, larger collagen fascicles in the lesions. Fibroblasts cultured from normal dermis, hypertrophic scar, and keloid for 5–6 weeks were intensely stained for FN. Extracellular matrix was heavily positive in cultures from the lesions compared with those from normal dermis.

Microvasculature in hypertrophic scars and the effects of pressure.

The fine structure of the microvasculature was compared among eight samples of normal skin, 79 granulation tissues, 48 hypertrophic scars, 11 hypertrophic scars treated with mechanical pressure, and 13 mature scars. Increased synthesis activity is suggested in endothelial cells from granulation tissues, is less in hypertrophic scars, and low in mature scars. In hypertrophic scars most of the microvessels appear partially or completely occluded. Endothelial cell nuclei are crenated, many villous projections from the endothelial cell membranes exist on the blood side, and endothelial cell junctions are often complex, although no large gaps are observed. In all the granulation tissues studied fibrin polymer is present, occurring intraluminally and interstitially, which may be related to endothelial cell proliferation. Therapeutic mechanical pressure over 1 to 3 months effects striking changes in endothelial and perivascular satellite cells. Rented areas appear in endothelial cell cytoplasm. A few such areas were found in cases of nontreated hypertrophic scars but in no other group. Pressure-treated scars also demonstrate degenerating perivascular satellite cells, which also are observed in a few cases of mature scars but in no other group. A previously published theory that hypoxia is related to generation of the hypertrophic scar, and that pressure probably increases hypoxia, resulting in long-term focal degeneration of selective cells, appears further supported by the present findings.

Increased fibronectin production by cell lines from hypertrophic scar and keloid.

Primary cell lines of fibroblasts from 8 tissues were established--three from hypertrophic scars (HS), one keloid (K) and four from the normal uninvolved dermis adjacent to each lesion. The objective was to quantify and compare all eight cell lines on the basis of fibronectin (FN) produced per cell and per total protein (PR). Two hypertrophic scars and their adjacent skin cell lines were evaluated by the ELISA method for FN and a micro Lowry assay for PR. The scar lines showed statistically significant increases in the amount of FN/cell compared to the cell lines from their adjacent normal dermis. The third hypertrophic scar and the keloid with their adjacent skin cell lines were assayed for FN and PR by radioimmunoprecipitation. Subconfluent cells were metabolically labeled with 35S-methionine for 20 hours. Harvested media and cell monolayers were assayed for radioactivity incorporated into FN and PR. The percentage of FN/PR was significantly higher in media for HS and K compared to the adjacent normal skin lines in the three passages tested. These results support our previous immunofluorescence studies and demonstrate that a fibroblast-type cell line from a hypertrophic scar or keloid produces more FN/PR over time than the normal fibroblast-type cell line from adjacent uninvolved dermis.

Ultrastructure of the Vaginal Tissue of Rabbits Treated With Collagen Sponge Alone and Medicated with Zinc and Copper Salts and Copper Wire

We tested the reaction of rabbit vaginal epithelium to inserted collagen sponges which have been proposed for human use as intravaginal contraceptive barriers. Some collagen sponges were medicated with zinc and copper sulfate and oher (untreated) sponges were wrapped loosely with copper wire. Scanning and transmission electron microscopy was performed on vaginal tissue after 10days of intravaginal contact with the sponges. The characteristic longitudinal rugate arrangement of polygonal, flat epithelial cells with a dense, even covering of microvilli seen in intact vaginas was not changed by the presence of a collagen sponge for 10days. In the presence of zinc sulfate in the sponge, the microvilli were in general shorter; however, the cells remained flat and continuous, forming visible, longitudinally oriented folds. Sponges containing copper sulfate induced marked changes in the epithelial lining of the rabbit vagina. Cells of differing sizes protruded above the plane of the surface. These cells had a hypertrophic appearance with narrow neck and widening at the top. Microvilli in these cells were very short and widely spaced, in contrast to the profuse microvillous projections in other vaginal samples. Copper wire induced definite abnormalities which were classified into three categories: (1) altered appearance similar to that caused by copper sulfate; (2) localized complete denudation of cells; (3) necrotic changes with penetration of the continuity of epithelial lining and excessive leukocytic infiltration of the lesion at the site of direct contact of the vaginal mucosa with the wire.

The use of a transport medium (L15M15) for bulk tissue storage and retention of viability

Twenty-five human or mouse tissue samples, some up to 8×4×2 cm, were immersed in a special transport medium (TM), L15M15, up to 7 days before being processed or placed in tissue culture. To test the efficacy of this medium, we concurrently placed pieces of the same tissues in a sterile phosphate buffered solution (PBS). We also tested the preservative capabilities of TM and PBS at room temperature and with refrigeration. Differences between TM and PBS are demonstrated, which are more pronounced using room temperature up to 4 days time. The tissues stored in TM show fewer degenerative or autolytic changes than the same tissue stored in PBS under identical conditions. Using regrigeration further enhanced the preservative qualities of TM up to 4 days, but not PBS. There were no obvious differences between tissues stored in TM and PBS with refrigeration after 7 days. We conclude that transport medium L15M15 is a useful medium for preserving tissue viability, especially large tissue samples, up to 4 days, especially if refrigerated.

A New Wave Dialectic 1 The Reinvention of Human Embryology

In 1973 Roe v. Wade was adjudicated by the Supreme Court. This landmark decision proved to be the watershed between science and the law. Statements made within the decision, and since, concerning human development, have been disingenuous, irresponsible or deliberately deceitful. One would like to believe that Supreme Court Justices, acting as learned and wise servants of our society, exercise great and considerate care in making decisions that not only affect our daily lives, but impact the evolution of our culture in the most moral and responsible way. We also want to believe they seek out all available facts concerning a case before coming to decisions. Alas, such is not the case.

In Defense of Human Development

Efforts to influence policy decisions relative to abortion, whether on the federal or state level, have inevitably included statements about human development invoked especially to support a particular political point of view. The cumulative effect of the many statements has given rise to an ugly spectre of human development, which is being reinvented as a derivative of the sociolegal aspect of the abortion issue. Since Roe vs. Wade, in 1973, more public statements have been made concerning human development than perhaps in all of our previous recorded history. Many of those statements have been misguided, if not outright inaccurate. The abortion issue has crystallized the publics need to know the truth about our own development. Yet, the significance of this truth goes far beyond the abortion issue. Elective abortion is intervention. But, in a similar sense, so are in-vitro fertilization, in-utero fetal surgery, fetal tissue research, drug addicted newborns, smoking pregnant women (and proximal consorters) and pregnant women who drink and produce fetal alcohol syndrome babies. Because of the consequences to human development by the above-mentioned procedures and conditions, and with the advent of such technology as gene synthesis, selection, modification and repair, it is high time we take a closer look at our developmental history. Four major concepts have fallen prey to contemporary socio-Iegal issues: The Beginning of Life, The Quality of Being Human, Viability, and Sentience.

There Is No Such Cell as a Human Embryonic Stem Cell– At Least, Not Yet

Abstract Separate and apart from the scientific truths of the tenets of human embryology taught to all medical students by lecture and text, political parsing of these tenets has been rampant for many years. It is time to reclaim these scientific truths before it becomes too late. The American Association of Anatomists should allow for a fair and balanced dialogue on human embryonic stem-cell research. It further should accept the fact that a true human embryonic stem cell has not been identified—at least, not yet.

The Final Corruption of Human Embryology

Abstract In 1993 I warned that human embryology was being rewritten according to political correctness. My prediction has come true. In 2005 a new textbook, Bioethics and the New Embryology , by Scott F. Gilbert, Anna L. Tyler, and Emily J. Zacklin, was published. It is a monument to political correctness and a testimony to the final corruption of human embryology.