Pacific Hair

RECENT SCIENTIFIC WORK ON HAIR GROWTH, WORLD WIDE: Part 3

admin — Thu, 17 May 2012 08:39:02 +0000

13)   Dr. Klugluger of Vienna with the Mosier Clinic has found that a tissue culture medium containing inhibitors of nitric oxide has resulted in grafts with no transient hair loss that begin growing immediately after hair transplantation. A variety of additives have been included in storage solutions, most commonly energy substrates and anti-oxidants. DMEM-containing inhibitors of inducible nitric oxide synthase (iNOS) prevented post-transplant hair shedding in grafts in six or six patients. The primary inhibitor of nitric oxide synthase was amino guanidine. DMEM containing arachidonic acid inhibitors prevented graft hair shedding in five of six patients versus zero of six in controls. Both additives also demonstrated significant improvement in hair shaft elongation studies.
14)   Work is currently being conducted to determine the effectiveness of an inexpensive ATP supplementation preparation called Lipo-Tripolyphosphate, which would be applied topically postoperatively in hair transplantation.
15)   Postoperative anti-oxidants which could be administrated orally include vitamin C and vitamin K.
16)   Platelet-rich plasma contains growth factors such as PDGF, TGF Beta 1, and VEGF. Using this for grafts and donor healing may result in a 15% increase in growth.
17)   Grafts will grow better if left slightly chubby with a little tissue beyond the dermal sheath and papilla.
18)   Dr. Rinaldi of Milan studied the twice-daily topical application of a postop solution containing adenosine sulfate 0.1%, taurine 1.0%, and ornithine chloride 1.0% (called 1-3 Atodine). Adenosine sulfate regulates vascular endothelial growth factor (VEGF) and follicular growth factor-7 (FGF-7), while taurine and ornithine stimulate outer root sheath growth. At one month, vessel diameter and hair shaft diameter were both larger than placebo. Revascularization was quicker by nearly threefold, and the follicle growth was improved.
Since the outer root sheath is more accessible to topical therapy than the dermal papilla, it may be that topical 1-3 Atodine solution might be effective.

RECENT SCIENTIFIC WORK ON HAIR GROWTH, WORLD WIDE: Part 2

admin — Tue, 15 May 2012 15:02:14 +0000

7)   Similarly, topical T3 solution postop may also contribute to a shortening of the telogen phase or its elimination. Another recent report has suggested that there may be little or no type 2 5-alpha reductase located in occipital (donor) dermal papilla cells. Of course this would explain, if true, why occipital dermal papilla cells are relatively impervious to the effects of DHT, since most DHT is located intracellular and not in the circulation. It may be that there are very low levels of DHT in these cells due to the lack of 5-alpha reductase type 2. It may also be that testosterone, not DHT, is the stimulator for the growth factors in these cells.
8)   Dr. Perez- Mesa, et al. found that growth factor activity appeared as follows following hair transplantation:
TGFA appeared on day 1, others appeared on day 3. Growth factors increased on days 7 through 21. By day 28, EGF and BEGF were the only ones present.
Three phases of revascularization of hair grafts were seen:
1)   Plasmatic imbibition at days 1 to 3.
2)   Primary inosculation and growth of blood vessels.
3)   Secondary inosculation/neovascularization with changes in the vessels including the lymphatics. These changes took place simultaneously with the increase in growth factors.
The study showed that after the hair graft takes and survives, the process may continue with one of the following scenarios:
a)   The hair will continue to grow in anagen phase from day one with no hair loss.
b)   The hair will fall out from one to six weeks postop (anagen effluvium) with new hairs beginning to grow two to four weeks later.
c)   The hair will fall out from seven to twelve weeks postop (telogen effluvium) with new hairs beginning to grow two to four weeks later. Hairs with individual follicular units do not grow at the same rate and may take any of the above three alternatives. They conclude that maintaining a healthy blood supply to the scalp and minimizing hypoxia and ischemia of the hair graft during transplant surgery and for five days postop should increase survival rate.
9)   Dr. Cooley has found that ischemia reperfusion injury results in 600% increase in free radicals within the hair follicles. This can be decreased by:
1)   Giving the patient anti-oxidants such as vitamin E or melatonin.
2)   Corticosteroids.
3)   Additives to graft holding solutions. Hypothermasol contains two potent anti-oxidants, glutathione and a synthetic analog of vitamin E.
Dr. Cooley’s current practice is to keep a mini-fridge in the O.R. and placed unslivered strips in Hypothermasol there. Slivers are also kept in Hypothermasol on coolers. It is important to keep Hypothermasol cold, and they switch out coolers at lunch. In this way, we keep the solution at 10°C or less. Thus all tissue has been soaked in antioxidants for theoretical protection against ischemia reperfusion injury.
The dissected grafts are then kept on DMEM/HEPES culture media. DMEM storage media stands for Dullbecco’s Modified Eagles Medium. This culture media contains glucose, vitamins, and a buffer to keep the pH steady. He keeps the grafts on DMEM-soaked Reston foam at the air liquid interface as opposed to immersion. He does not chill them. Both the Hypothermasol and DMEM have the HEPES buffer in them which keeps the pH steady.
10)   Dr. Cotsarelis has found that stem cells in hair follicles present as keratinocytes, which maintain a high proliferative potential and are long lived, slowly cycling, and apparently immune privileged cells. Topical treatment with RU486 to these bulged stem cells resulted in hair follicles entering the anagen phase after only five days. These cells have the potential to regenerate new hair follicles at about four weeks after implantation into mouse skin.
11)   The regular spacing of hair follicles throughout the scalp and elsewhere is controlled by an Edar/BMP activation-inhibition mechanism, which operates along side a label pre-pattern, suggesting that Edar-mediated stabilization of beta-catenin active foci is a key event in determining definitive follicle locations. The BMPs repress epidermal Edar and hence, follicle rate. Edar activation also induces connective tissue growth factor, an inhibitor of BMP signaling, allowing BMP action only at a distance from their site of synthesis. Consistent with this model, transgenic hyperactivation of Edar signaling leads to widespread overproduction of hair follicles.
12)   Plasma-lyte A has a pH of 7.4 and uses an acetate buffer. DMEM used in hair studies normally contains the more expensive HEPES buffer, which works well in open-air situations.

RECENT SCIENTIFIC WORK ON HAIR GROWTH, WORLD WIDE: Part 1

admin — Mon, 14 May 2012 21:45:31 +0000

1)   Hormone replacement therapy is frequently used as a technique for anti-aging in Southern California and other locations. People are usually prescribed various vitamins, supplements, thyroid hormone (in the form of Armour thyroid, which is T3 and T4), growth hormone, and sex hormones such as Testosterone or Estradiol. The theory goes that as we age, the ability to convert T4 into T3 (which is done in the tissues) diminishes so that persons over 55 are frequently deficient in free T3, even though they may have sufficient T4.
2)   It has been observed for several years that grey hair tends to darken up on these therapies. It was previously speculated that this was largely due to the growth hormone. The article by Redondo, et al., in the Forum, March/April 2008, shows two mice whose hair darkens up on thyroid hormone. More interestingly, the follicles of the “thyroid solution stimulated mouse” entered Anagen phase at day 6 compared to day 10 or 16 for the mouse without thyroid solution applied. Similar results were found with follicular units in a thyroid solution. The thyroid solution is tri iodothyronine (T3).
3)   About five years ago, the author transplanted a 50-year-old man, who postoperatively had relatively sparse results from his hair transplant. After his hair transplant, his donor hair began to thin, so that some sort of diffuse thinning process was suspected. A hormonal workup was advised, but he declined at that time. A year or two later, he had a hormonal workup from his internist, who then put him on AndroGel (testosterone). Within a few months of being on AndroGel, the patient reported that all of his hair, both transplanted and original donor hair, thickened up and developed a luxurious quality that he had had when he was much younger.
We have known for a long time that body hair and beard hair requires the support of testosterone. We have also all assumed that scalp hair is negatively affected by DHT.
Based on the above observations in this patient, it would appear that donor hair requires testosterone to support it, as well as body hair and beard hair. This may explain why some late-middle-aged men have diffuse thinning of their hair. Perhaps all should have an endocrine panel and then be treated with testosterone, as well as T3, and other hormones as indicated by lab evaluation.
4)   Another observation by Dr. Neal Rouzier in Palm Springs, CA, who has a large anti-aging practice, relates the two following facts. 1) A very common cause of depression in postmenopausal women is low free T3. Women who are at the low end of the normal range are frequently treated by Dr. Rouzier to bring them up to the high end of the normal range of free T3 or even a little higher. By titrating free T3, he is able to reverse the depression, reverse the loss of energy, observe darkening of white hair, and thickening of hair overall. He also has communicated to me that generally postmenopausal women have immeasurable levels of androgens. That being the case, perhaps some of the diffuse hair thinning commonly seen in postmenopausal women is related to loss of androgen support for their donor hair. Perhaps treatment with testosterone for both postmenopausal women and for aging men will be a principal factor, along with thyroid hormone, in reversing the DPA or diffuse thinning situations that are commonly seen.
5)   In looking at the article on gene expression by Drs. Kim and Kim of Korea in the July/August Forum, they observed that DHT in frontal cells of the scalp induced DKK-1, which inhibits hair growth. Contrarily, in beard dermal papilla cells, DHT stimulated the EDA pathway which caused hair growth of the beard cells. Is it likely that occipital and parietal donor area cells are stimulated by DHT (and/or testosterone) to activate the EDA pathway, rather than the DKK-1 pathway? It would seem that this is probable, based on the observations.
In general, activation of the Wnt pathway in hair follicles and insulin-like growth factors (IGFs) are involved in androgen induction, and TGF-β pathways are related to catagen progression. Frontal DP cells also more predominantly produced many stress proteins including heat shock proteins. Thus, frontal DP cells are, before androgen stimulation, more severely stressed cells.
DKK-1 is induced by DHT in frontal DP cells and inhibits hair growth.
Ectodysplasin A (EDA) pathway is activated in beard DP cells by DHT stimulation and induces anagen hair growth.
In beard DP cells, DKK-1 was not induced by DHT, as it was in frontal balding DP cells. DHT rapidly increased the EDA2R pathway stimulating hair growth in beard DP cells.
6)   Nilofer Farjo writes in her “Hair Sciences” column in the March/April issue of the Forum that the addition of IGF-1 or 2 (insulin growth factor) maintains antigen growth in vitro in dermal papilla cells. IGF-1 is essentially growth hormone, and so the question presents itself whether or not using perhaps a double dose of growth hormone on patients for two or three months after a hair transplant would get the hairs growing a lot sooner. Perhaps it would eliminate the hair shock and three months of telogen, which is commonly experienced. That therapy could be systemic or topical. Research will need to be done with this.

In Vitro Growth of Hair Follicle Cells

admin — Sun, 13 May 2012 20:56:51 +0000

Mass-Szabowski et al., (2001), reported that organotypic cocultures of keratinocytes and fibroblasts generate normal epithelial cells. These results were obtained with fibroblasts from any species of any tissue type. Further, they report that the “use of mouse fibroblasts and human keratinocytes facilitates the identification of the origin of compounds involved in epidermal tissue reconstitution and growth regulation. Moreover, the functional significance for the keratinocyte phenotype of genetically modified fibroblasts from transgenic or knockout mice, even those exhibiting an embryonic lethal phenotype, can be studied in such heterologous in vitro tissue equivalents. Here we communicate results of such studies revealing the antagonistic function of mouse fibroblasts defective in the AP-1 constituents c-June and JunB, respectively, on human keratinocyte growth and differentiation. Furthermore, the hematopoietic growth factor granulocyte macrophage-colony stimulating factor has been identified as a novel regulator of keratinocyte growth and differentiation. As will be reported in detail elsewhere, both granulocyte macrophage-colony stimulating factor and keratinocyte growth factor have been identified as major mediators of fibroblast-keratinocyte interactions and their expression is induced via AP-1 by interleukin-1 released by the epithelial cells. Thus, these heterologous cocultures provide a novel promising tool for elucidating molecular mechanisms of epithelial-mesenchymal interactions and their consequences on epithelial cell proliferation and differentiation.”

Sonic Hedgehog: Part 3

admin — Sat, 12 May 2012 21:11:05 +0000

Martin from Madrid has the following report, “The Hedgehog (Hh) family of signaling molecules function as organizers in many morphogenetic processes. Hh signaling requires cholesterol in both signal-generating and –receiving cells, and it requires the tumor suppressor Patched (Ptc) in receiving cells in which it plays a negative role. Ptc both blocks the Hh pathway and limits the spread of Hh. Sequence analysis suggests that it has 12 transmembrane segments, 5 of which are homologous to a conserved region that has been identified in several proteins involved in cholesterol homeostasis and has been designated the sterol-sensing domain (SSD). In the present study, we show that a Ptc mutant with a single amino acid substitution in the SSD induces target gene activation in a ligand-independent manner. This mutant Ptc(SSD) protein shows dominant-negative activity in blocking Hh signaling by preventing the downregulation of Smoothened (Smo), a positive effector of the Hh pathway. Despite its dominant-negative activity, the mutant Ptc protein functioned like the wild-type protein in sequestering and internalizing Hh. In addition, we show that Ptc(SSD) preferentially accumulates in endosomes of the endocytic compartment. All these results suggest a role of the SSD of Ptc in mediating the vesicular trafficking of Ptc to regulate Smo activity.” Strutt et al., MRC Intercellular Signaling Group, Centre for Developmental Genetics, Department of Biomedical Science, University of Sheffield, Sheffield, United Kingdom, “The tumor suppressor gene patched (ptc) encodes an approximately 140 kDa polytopic transmembrane protein that binds members of the Hedgehog (Hh) family of signaling proteins and regulates the activity of Smoothened (Smo), a G protein-coupled receptor-like protein essential for Hh signal transduction. Ptc contains a sterol-sensing domain (SSD), a motif found in proteins implicated in the intracellular trafficking of cholesterol, and/or other cargoes. Cholesterol plays a critical role in Hedgehog (Hh) signaling by facilitating the regulated secretion and sequestration of the Hh protein, to which it is covalently coupled. In addition, cholesterol synthesis inhibitors block the ability of cells to respond to Hh, and this finding points to an additional requirement for the lipid in regulating downstream components of the Hh signaling pathway. Although the SSD of Ptc has been linked to both the sequestration of, an the cellular response to Hh, definitive evidence for its function has so far been lacking. Here we describe the identification and characterization of two missense mutations in the SSD of Drosophila Ptc; strikingly, while both mutations abolish Smo repression, neither affects the ability of Ptc to interact with Hh. We speculate that Ptc may control Smo activity by regulating an intracellular trafficking process dependent upon the integrity of the SSD.

Sonic Hedgehog: Part 2

admin — Sat, 12 May 2012 08:05:41 +0000

In a report entitled “Hedgehog signal transduction: from flies to vertebrates,” by Murone et al., it was recently shown that the patterning and morphogenesis of multicellular organisms require a complex interplay of inductive signals which control proliferation, growth arrest, and differentiation of different cell types. A number of such signaling molecules have been identified in vertebrates and invertebrates. The molecular dissection of these pathways demonstrated that in vertebrates, mutations or abnormal function of these signaling pathways were often associated with developmental disorders and cancer formation. The Hedgehog (Hh) family of secreted proteins provides a perfect example of such signaling proteins. In the following review, we will not discuss in detail the role of Hh as a morphogen, but rather focus on its signal transduction pathway and its role in various human disorders.
The Hedgehog induces opposite changes in turnover and subcellular localization of patched and smoothened. It was demonstrated by Denef et al., (2001), from Heidelberg, Germany, that secreted signaling proteins of the Hedgehog family organize spatial pattern during animal development. Two integral membrane proteins have been identified with distinct roles in Hedgehog signaling. Patched functions in Hedgehog binding, and Smoothened functions in transducing the signal. Current models view Patched and Smoothened as a preformed receptor complex that is activated by Hedgehog binding. Here we present evidence that Patched destabilizes Smoothened in the absence of Hedgehog. Hedgehog binding causes removal of Patched from the cell surface. In contrast, Hedgehog causes phosphorylation, stabilization, and accumulation of Smoothened at the cell surface. These findings raise the possibility that Patched acts indirectly to regulate Smoothened activity.
Kalderon has reported that the sonic Hedgehog receptor patched associates with caveolin-1 in cholesterol-rich microdomains of the plasma membrane and Karpen et al., (2001), reported that the Hedgehog signaling pathway is involved in early embryonic patterning as well as in cancer; however, little is known about the subcellular localization of the Hedgehog receptor complex of Patched and Smoothened. Since Hh has been found in lipid rafts in Drosophila, we hypothesized that Patched and Smoothened might also be found in these cholesterol-rich microdomains. In this study, we demonstrate that both Smoothened and Patched are in caveolin-1-enriched/raft microdomains. Immunoprecipitation studies show that Patched specifically interacts with caveoloin-1, whereas Smoothened does not. Fractionation studies show that Patched and caveolin-1 can be co-isolated from buoyant density fractions that represent caveolae/raft microdomains and that Patched and caveolin-1 co-localize by confocal microscopy. Glutathione S-transferase fusion protein experiments show that the interaction between Patched and caveolin-1 involves the caveolin-1 scaffolding domain and a Patched consensus binding site. Immunochemistry data and fractionation studies also show that Patched seems to be required for transport of Smoothened to the membrane. Depletion of plasmalemmal cholesterol influences the distribution of the Hh receptor complex in the caveolin-enriched/raft microdomains. These data suggest that caveolin-1 may be integral for sequestering the Hh receptor complex in these caveolin-enriched microdomains, which act as a scaffold for the interactions with the Hh protein.

More Before & After Photos

admin — Fri, 11 May 2012 06:26:29 +0000

Sonic Hedgehog: Part 1

admin — Wed, 09 May 2012 15:16:27 +0000

Hooper is currently doing research at the University of Colorado on two secreted proteins, Wingless and Hedgehog. Wingless (Wg) is expressed by the most posterior cells in each parasegment; Hedgehog (Hh) is expressed in the most anterior cells of the next parasegment. Immediately after gastrulation, the two cell types are mutually dependent. Local Wg signaling stabilized Hh expression and local Hh signaling stabilizes Wg expression. Direct Wg autoregulation (autocrine signaling) is masked by its paracrine role in maintaining Hh, which in turn maintains Wg. I have used zeste-white 3 (zw3) and patched (ptc) mutant backgrounds to uncouple genetically this positive-feedback loop and to study autocrine Wg signaling. I report here that direct Wg autoregulation differs from Wg signaling to adjacent cells in the importance of fused (fu), smoothened (smo) and cubitus interruptus (ci) relative to zw3 and armadillo (arm). I also find that Wg autoregulation during this early Hh-dependent phase differs from later Wg autoregulation by lack of gooseberry (gsb) participation.
Zhu et al., (1999), has stated that although Sonic Hedgehog (Shh) plays a critical role in brain development, its actions on neural progenitor cell proliferation and differentiation have not been clearly defined. Transcripts for the putative Shh-receptor genes patched (Ptc) and smoothened (Smo) are expressed by embryonic, postnatal, and adult progenitor cells, suggesting that Shh can act directly on these cells. The recombinant human amino-terminal fragment of Shh protein (Shh-N) alone did not support the survival of cultured progenitor cells, but treatment with Shh-N in the presence of bFGF increased progenitor cell proliferation. Furthermore, treatment of embryonic rat progenitor cells propagated either in primary culture or after mitogen expansion significantly increased the proportions of both beta-tubulin- (neuronal marker) and O4- (oligodendroglial marker) immunoreactive cells and reduced the proportion of nestin- (uncommitted neural progenitor cell marker) immunoreactive cells. By contrast, Shh-N had no effect on the elaboration of GFAP – (astroglial marker) immunoreactive cells. Cotreatment with Shh-N and bone morphogenetic protein-2 (BMP2) inhibited the anti-proliferative, astroglial-inductive, and oligodendroglial-suppressive effects of BMP2. Our observations suggest that Shh-N selectively promotes the elaboration of both neuronal and oligodendroglial lineage species and inhibits the effects of BMP2 on progenitor cell proliferation and astroglial differentiation.

Transfecting of Human Hair Follicle Cells

admin — Sat, 05 May 2012 21:59:42 +0000

Gupta el al., (2001), reported that the human hair follicle consists of plastic cells for hair follicle cycling and for epidermal keratinocytes, melanocytes, and Langerhans cells. They found that a combination of liposomes and vector DNA can be used to target hair follicle cells in human scalp xenografts. The liposome composition and stage of the hair cycle were found to be important parameters influencing transfection of human hair follicles. Transfection was only successful during anagen onset. Hoffman (1998) has reported that liposomes may also be used topically to deliver genes to hair follicles.
Conclusion
It is important to have cells which survive long enough to permit us to study their properties and which signal molecules control their differentiation. Adult human hair follicle cells do not proliferate and die in culture after several weeks. After transfection with hTERT, we now have the ability to increase their lifespan and study their differentiation under the control of signal peptides.
It is not clear how hair follicle morphogenesis is controlled, but it appears that many signal proteins are involved. Mesenchymal and epithelial cells use fibroblast growth factor and bone morphogenic proteins to provide the critical signals to the stem cells.
The use of mineral ascorbates in modern medicine has the ability to improve many of the above reviewed processes. It is imperative the researchers work together and use mineral ascorbates in every procedure to validate their beneficial effects. Every cell process requires mineral ascorbates in one way or another. It is the job of the research community to serve the consumer and devote substantial research efforts to the mineral ascorbates which the public has already started to consume in great quantities.

T3 – Thyroid Hormone

admin — Fri, 04 May 2012 21:24:59 +0000

Hormone replacement therapy is frequently used as a technique for anti-aging in Southern California and other locations. People are usually prescribed various vitamins, supplements, thyroid hormone (in the form of Armour thyroid, which is T3 and T4), growth hormone, and sex hormones such as Testosterone or Estradiol. The theory goes that as we age, the ability to convert T4 into T3 (which is done in the tissues) diminishes so that persons over 55 are frequently deficient in free T3, even though they may have sufficient T4.

It has been observed for several years that grey hair tends to darken up on these therapies. It was previously speculated that this was largely due to the growth hormone. The article by Redondo, et al., in the Forum, March/April 2008, shows two mice whose hair darkens up on thyroid hormone. More interestingly, the follicles of the “thyroid solution stimulated mouse” entered Anagen phase at day 6 compared to day 10 or 16 for the mouse without thyroid solution applied. Similar results were found with follicular units in a thyroid solution. The thyroid solution is tri iodothyronine (T3).