Titanium alloy (Ti-6Al-4V) has been widely used in medical field due to its good biocompatibility and machinability. However, Ti-6Al-4V lacks antibacterial ability, which can be easy to lead the surgical infection by bacteria. At the same time, the release of the toxic ions such as Al or V in Ti-6Al-4V to the human body under the physiological environment is also easy to cause harm to the human body. Therefore, in order to solve the above problems, this study first used magnetron sputtering technique to prepare TiN-Cu composite coatings on the surface of Ti-6Al-4V, which is expected to take advantage of Cu antibacterial properties to achieve long-term antibacterial effects while preventing the release of toxic ions. Second, in order to enhance cell adhesion of the TiN-Cu composite coatings, plasma immersion ion implantation (PIII) was used to implant Mg ions into the surface of the composite coatings respectively. A series of measurements (such as XRD, XPS, SEM, etc.) were used to analyze the structure, composition and mechanical properties of TiN-Cu composite coatings before and after doping with different ion doses. L929 cells and MC3T3-E1 cells were used to analyze the cell proliferation and adhesion on the sample surfaces. The survival of Escherichia coli by using the coating plate method. The results showed that the surface of Mg⁺ implanted TiN-Cu composite film had the best antibacterial property and cell proliferation ability with the highest protein adsorption ability. Cu trapped electrons on the bacteria's surface and destroyed their membranes. In addition, the bonding of Cu and respiratory enzymes in the bacteria also caused bacterial dysfunction. The implanted Mg⁺ on the surface stimulated protein adsorption, the cell adhesion and proliferation.

The contact angle method [1] quickly and noninvasively evaluates skin wettability with water to determine the skin hydration state [2]. Athletes' body hydration state influences their skills and performance, affecting the outcome of a game [3].

Our objective was to measure in vivo the changes in skin hydration of 12 basketball boys (age 13- to 15 years) before and after (called here “response”) a game about basal hydration conditions (called here “normal conditions”) and after oral assumption of mineral water 2L per week (called here “hydro conditions”) before a second game. Basal and response skin hydration measurements were performed on the right (dx) and left (sx) forearms skin. The response measurements were done 10’ after the end of the game and on the skin without sweat.

The skin hydration state was assessed using the contact angle (CA; deg) method with a mobile Tenskinmeter®. Personalized evaluations of epidermal moisture changes were conducted utilizing the Skin Hydration Debt (SHD) parameter, introduced by Davide Rossi in 2018, following the “Basketskin protocol”. A positive SHD indicates that the skin is experiencing “water debt,” whereas a negative value suggests “water credit”. The initial findings revealed a noticeable increase in skin hydration state after the first game, which correlated with a decrease in the average water contact angles observed at the interface of the right (dx) forearm skin (N=24) (basal CA: 106.7±18.3 deg, response CA: 78.8±21.1 deg) and left (sx) (N=24) forearm skin (basal CA: 111.5±15.7 deg, response CA: 79.0±17.0 deg). 

As regards the hydro conditions, the results showed a trend increase in skin hydration state before and after the second game corresponding to a trend decrease of droplet average water CAs at the interface with dx (N=24) forearm skin (basal CA: 104.0±17.7 deg, response CA: 62.7±20.1 deg) and sx (N=24) forearm skin (basal CA: 103.8±20.5 deg, response CA: 62.4±18.0 deg). 

The negative difference (Δ) between the basal (b) average CAs absolute values (|CAs|) measurements performed in hydro (h) conditions and that performed in normal (n) conditions on dx and sx forearms and the negative difference between the average response (r) average CAs absolute values (|CAs|) measurements performed in hydro (h) conditions and that performed in normal (n) conditions on dx and sx forearms were respectively Δ|CA|_b^dx=2.65 deg, Δ|CA|_r^dx=16.1 deg, Δ|CA|_b^sx=7.68 deg, and Δ|CA|_r^sx=16.45 deg. 

These data demonstrated that 2L water administration per week causes an overall a trend decrease of skin wettability with water drop, however the major trend increase of hydration state was revealed from the CAs measured after the two games (responses).

The second result demonstrated the capability of Skin Hydration Debt (SHD) in the evaluation of the hydration state of each boy before and after the two games, and in “normal” and “hydro” conditions. In the case of normal conditions (first game), basal average SHD related at dx  forearm skin (basal SHD: -1.15±8.00, response SHD: -11.45±13.7) and left (sx) forearm skin (basal SHD: +1.23±5.00, response SHD: -10.20±9.2) showed average water debt only in the case of basal (sx) forearms. For all other cases, the skins demonstrated be in “water credit”.  As regard hydro conditions (second game), basal average SHD related at dx forearm skin (basal SHD: -1.22±8.4, response SHD: -21.73±14.4) and left (sx) forearm skin (basal SHD: -1.36±8,4, response SHD: -21.76±12.5) demonstrated an increase “water credit” respect to “normal condition”

The difference (Δ) between the average |SHD| values of basal (b) and response (r) in hydro and normal conditions are respectively Δ^b(dx) _|SHD|=0.08, Δ^r(dx)_|SHD|=10.3, Δ^b(sx) _|SHD|= 0.14, and Δ^r(sx)_|SHD|=11.6.

Results demonstrated that the CA method is capable to determining in non-invasive and rapid way the real effect of water oral administration on the skin hydration state of basketball boys before and after the game. 

The SHD demonstrated its usefulness in the evaluation of the increase in skin hydration followed by the oral water administration performed by basketball boys one week before the second game mostly in the response evaluation.

This appears in accordance with the increase of body hydration following one week of water oral administration and improvement of skin moisturize that seems to promote the migration of water from the body inside to outside during the game. 

Our work is open to the possibility of using the tenskinmeterâ for control of the hydration state of sportsmen in relation to their real body hydration before the game.

The results open new perspectives in a large-scale application of the CA method to evaluate the increase of body hydration after oral assumption of water added with carbohydrate-electrolyte solution (CES) [3] and perform deep statistical analysis.

The surface tensiometry technique leads to determine skin hydration status [1], using the contact angle method [2] with water as the liquid test, and surface tension of a liquid,  using pendant drop method [3, 4], in a quick and non-invasive way. Bioenergy Field Treatments represent an approach to treating chronic diseases by assessing an individual's physiological and emotional responses through their own bioenergetic body, and is an integration into traditional medicine [5]. Our first objective was to measure the bio-informational fields applied at mineral and sparkling water drops to evaluate differences in their volume. 

Our second objective was to provide the bio-informational field treatment with a tool capable of measuring effectiveness through analytics capable of determining changes in skin hydration levels and surface tensiometry. Two Prano Surface Tensiometry Units were implemented in Milan and Maserada Sul Piave (Italy) to realize our research aims.

The bio informational fields were applied in vivo by placing the hands at a 5 cm distance for 3' from the test subject's skin and pendant natural and sparkling water drops [response X, Xs]. An expert pranopractic (true before [O] and after [X] treatment) applied the bio informational fields and a non pranopratic (sham before [Os] and after [Xs] treatment) as reference which responses were respectively [X] and [Xs]. The data were compared with correspondent controls [O] and [Os].

The results demonstrated that the effect of the action of true and sham pranopractics on the pendant drop volume variations have statistical differences (P-value = 0.00000 for both kind of waters for [O], [X] and [O,X], while for [Os] natural with [15]=23.32 and sparkling with t[15]=39.48 waters showed both p (>|t|) <.0001, [Xs] natural with t[15]=-68.75 and sparkling with t[15]=-77.95 waters showed both p (>|t|) <.0001, and for [Os, Xs] natural with t[15]=-7.90 and sparkling with t[15]=18.58 waters showed both p (>|t|) <.0.0001). 

The effect of the action of true and sham pranopractics on the water contact angle variations measured on forehead skin demonstrated statistical differences confirming that skin hydration change after the application of bio-informational fields by the true pranopractic to each test subjects involved in PTSU2 (p (>|t|) <.0001* for sham with t[8]=25.82 and true with t[8]=19.77 pranopractic both, p (>|t|)=0.6886 for sham with t[8]=-0.41 and p (>|t|) =0.8599 for true with  t[8]=-0.18 pranopractics). The analysis of variance of water contact angles measured in PTSU 2 also showed 95% subject CI [4.1013633, 163.12], 95% residue CI [23.08, 107.13], and p (Wald)=0.0303 confirming both the existence of trend variations of water contact angle between the true and sham pranopractics however the differences are not statistically significant. 

Results open the hypothesis of skin hydration variations trends and water pendant drop volume concerning control measurements after the bio informational fields application.

Our work combined non-invasive bio-informational field treatment with non-invasive surface tensiometry analytical approach using the contact angle method and water as a biocompatible liquid test. The results are preliminary and open new perspectives in a large-scale bio informational fields application to perform deep statistical analysis.

The measurement of contact angle (CA) on skin allows a simple and rapid test on the different pharmaceutical and cosmetic compounds [1]. Several studies evaluated the wettability of the skin surface as a function of the diffusion processes of the active ingredient through the skin [2]. The CA method can also be applied to evaluate skin hydration (SH) that is essential for body thermoregulation. Various investigations demonstrated correlations between water intake and variations in the SH status using the corneometric (H) approach and the variation in the Transepidermal Water Loss (TEWL) degree, respectively [3, 4], highlighting the influence of the intake of 2L per day on the SH. Our work aimed the assessing SH status by measuring the water CA and H, evaluating the influence of water intake and urination on SH status over time under controlled conditions involving four subjects (S1, S2, S3, and S4) aged between 24 and 26 between at environmental temperature of 24°C±0.5. 

As example, (a) S1 demonstrated a basal CA (°) of 83.3°, after 500 ml intake 75.4°(t₀), 65.4°(t₁₀), 82.4°(t₂₀), 80.1°(t₃₀), 88.4°(t₄₀), after urination (-200 ml) is 83.2°(t₀) and 83.7°(t₁₀), after 500 ml intake 77.6°(t₀), 76.6°(t₁₀), 79.4°(t₂₀), 77.7°(t₃₀), 79.9°(t₄₀), after urination (-300 ml) is 84.9°(t₀) and 91.9°(t₁₀), after 500 ml intake is 74.9°(t₀), 87.2°(t₁₀), 72,6°(t₂₀), 75.8°(t₃₀), 77.2°(t₄₀), after urination (-475 ml) is 67.0°(t₀) and 80.3°(t₁₀), (b) S2 demonstrated a basal CA (°) of 83.6°, after 500 ml intake 72.9°(t₀), 72.6°(t₁₀), 84.7°(t₂₀), 86.9°(t₃₀), 89.5°(t₄₀), after urination (-220 ml) is 85.4°(t₀) and 89.9°(t₁₀), after 500 ml intake 85.8°(t₀), 76.6°(t₁₀), 89.9°(t₂₀), 94.5°(t₃₀), 91.5°(t₄₀), after urination (-470 ml) is 96.7°(t₀) and 87.4°(t₁₀), after 500 ml intake is 89.0°(t₀), 83.1°(t₁₀), 90,6°(t₂₀), 92.5°(t₃₀), 91.2°(t₄₀), after urination (-400 ml) is 91.1°(t₀) and 88.4°(t₁₀), (c) S3 demonstrated a basal CA (°) of 83.8°, after 500 ml intake 78.1°(t₀), 83.0°(t₁₀), 84.4°(t₂₀), 84.2°(t₃₀), 88.3°(t₄₀), after urination (-210 ml) is 79.8°(t₀) and 82.9°(t₁₀), after 500 ml intake 82.1°(t₀), 82.6°(t₁₀), 79.1°(t₂₀), 84.4°(t₃₀), 83.2°(t₄₀), after urination (-580 ml) is 81.9°(t₀) and 84.9°(t₁₀), after 500 ml intake is 75.4°(t₀), 79.5°(t₁₀), 79,6°(t₂₀), 78.4°(t₃₀), 79.5°(t₄₀), after urination (-470 ml) is 85.9°(t₀) and 84.9°(t₁₀), and (d) demonstrated a basal CA (°) of 88.6°, after 500 ml intake 84.5°(t₀), 93.3°(t₁₀), 87.9°(t₂₀), 89.7°(t₃₀), 80.4°(t₄₀), after urination (-450 ml) is 78.3°(t₀) and 85.2°(t₁₀), after 500 ml intake 87.6°(t₀), 86.8°(t₁₀), 89.0°(t₂₀), 85.2°(t₃₀), 83.0°(t₄₀), after urination (-420 ml) is 86.7°(t₀) and 94.9°(t₁₀), after 500 ml intake is 85.9°(t₀), 87.8°(t₁₀), 89,5°(t₂₀), 85.9°(t₃₀), 84.5°(t₄₀), after urination (-500 ml) is 85.4°(t₀) and 86.6°(t₁₀). 

S1 showed a total average Hydration Index (HI) of 52.5±4.07 (Meas.1), 54.3±5.02 (Meas.2), 55.2±5.3 (Meas.3), S2 showed a total average HI of 35.1±1 (Meas.1), 37.2±5.1 (Meas.2), 37.2±4.9 (Meas.3), S3 showed a total average HI of 32.7±5.9 (Meas.1), 33.9±7.0 (Meas.2), 34.2±5.7 (Meas.3), and S4 showed a total average HI of 67.8±6.6 (Meas.1), 65.7±6.3 (Meas.2), 61.0±5.9 (Meas.3). In S1, the water intake caused increase in SH and decrease water CA. In the 1° period, CAs increase until the moment of urination because linked to the rapid hydrating effect due to the 1° water intake, and this phenomenon repeats at 2° water intake. 

In S2 the 1° and 2° water intake follow the same behaviour of S1, however the influence of water intake on SH appears more evident in S2 than in subject S1 because the difference between the volume ingested and excreted for the first two periods considered (1 Δ=280 mL, 2 Δ=30 mL) appears lower than that observed in S1 (1 Δ=300 mL, 2 Δ=200 mL). 

In S3, the 1° and 3° intake of water follow the same behaviour as S1 and S2, while after the 2° intake, the CAs showed less variability than what was observed in S1 and S2 in period 2. 

The S4 presents an anomalous CAs trend with respect to S1, S2 and S3 due to a poor ability to maintain body hydration levels over time and doesn’t appear suitable for the in vivo absorption test of a drug because it is closely linked to the maintenance of the SH status over time. 

Our work demonstrated that the CA method is capable of determining the influence of repeated water intake on the SH in relation to urination by measuring the CAs of a water droplet at different times. 

Our results open new perspectives in the evaluation of the effect of SH on the in vivo absorption of an active ingredient, developing a correlation model between CAs data obtained from static conditions and those obtained under kinetic conditions after application of the formulation loaded with the active ingredient.