2006 TRI/PRINCETON CONFERENCE Ingredient (INCI name) Water Hexapeptide-11 Fruit acids Betaine Green tea polyphenols Hexylene glycol IN VIVO TESTING PROTOCOL Table I Treatment Ingredients Percentage in composition 46-50 1-2 25-30 8-12 2-3 8-12 371 The entire study was conducted at a clinically-sponsored salon which the participants visited every day for five days. Prior to commencing the study, the participants signed informed consent agreements. Participants were all Caucasian women with a mixed population of damaged and non-damaged hair, between the ages of 30 and 60, of general good health and non-pregnant. Each participant was required to have hair of at least 12 inches in length. None of the participants reported signs of female alopecia or other known scalp conditions which might skew the test results. Prior to commencing the study, the participants went through a five day wash out period on their hair using a simple ionic shampoo and the placebo conditioner. In addition, after the wash-out period, but prior to the treatment period, each participant had a fabric collar transfixed around their neck so that during drying, any hairs which fell from the head were captured in the collar. The participant's hair on one half of their head (to remain consistent with the test results from the half-head study described below) was then dried for three minutes with a brush and a 1200 W hair dryer. At the end of the drying period, the clinician then gathered all of the hair fibers that were trapped in the brush and those that had fallen into the collar and these hair fibers were culled into plastic bags and labeled as "Day O." Only fibers that could be visibly seen were gathered (typically greater than ½-inch in length or greater). The participants came into the salon every day at the same time for their hair treatments. During the treatment, each participant was shampooed with the same simple shampoo and then the hair was combed to afford half-head treatment sites. To one side of the head, the salon clinician applied the placebo conditioner and allowed the conditioner to reside on the head for five minutes prior to rinsing. To the other side of the head, the participants were treated with the conditioner containing the active ingredients which were also allowed to reside on the scalp and hair for five minutes prior to rinsing. Each side of the hair was rinsed in such a way as to prevent treatment from one side of the head to wash over to the other side of the head. Such treatments were continued once a day for five complete days. At completion of the study period, after the final conditioning rinse, the participants were affixed with the same collar mentioned above and each side of the head was dried for three minutes. After completion of one side of the head, all the hair fibers were gathered before the other side of the head was similarly dried. The hair fibers were culled as before and placed into plastic Ziploc bags and labeled as "Day 5" with identification of whether they were from the active- or placebo-treated side of the head.

372 JOURNAL OF COSMETIC SCIENCE SEPARATION OF HAIR FIBERS The hair fibers that were gathered at Day O and Day 5 were transferred to a laboratory housing a stereomicroscope. The contents of each plastic bag were carefully examined in order to determine whether or not the hair contained an intact hair bulb. Care had to be taken here as some telogen hair fibers can look very much like broken hair fibers (i.e., the hair bulb is tiny). However, clinicians were able to meticulously separate hairs that had broken from hairs that were removed with intact hair bulbs. RES UL TS AND DISCUSSION Shown below are two SEM images of typical hair fibers in which the hair was removed from the scalp with an intact follicle bulb (Figure 1). As can be seen, the appearance of the hair fiber in telogen phase shows a very tiny hair bulb which can be mistaken for a broken hair. Broken hair fibers are generally quite easy to distinguish as are hair fibers that retain an anagen hair bulb. None-the-less, separation of the hair fibers is a tedious process which must be done carefully to assure accurate identification of the hair fibers as either intact or broken. After the hairs were separated the numbers of hairs in each group, Day O Intact, Day 0 Broken, Day 5 Treated-Intact, Day 5 Treated-Broken, Day 5 Placebo-Intact and Day 5 Placebo-Broken, were counted. The results of this study are shown in Figure 2. What becomes immediately apparent in looking at the data in Figure 2 is that in every instance, the number of hairs that were extracted intact from the scalp far exceeded the number of hairs that broke (by nearly a factor of six in every case). This result implies that the concept of hair loss must seriously take into account the difference between hairs that break (which we prefer to call brittleness) and those that are removed (extracted) fully intact from the scalp. This data also suggests that hair fiber strength studies which employ tresses and hair elongation techniques are overlooking a major route of hair removal which cannot, by definition, be accounted for in such ex vivo type studies. In addition, as these results are taken from a realistic treatment protocol, the mode of hair breakage, i.e., elongation versus hair entanglement on the brush tangs, is minimized. Figure 1. Scanning electron phoromicrographs of two intact hair fibers, one that is in apparent anagen phase growth (left) and the other that appears to be in telogen phase growth (right).